Dharam Paul Chaudhary

Sandeep Kumar Sapna Langyan

Editors

Maize: Nutrition
Dynamics and
Novel Uses
Maize: Nutrition Dynamics
and Novel Uses
Dharam Paul Chaudhary Sandeep Kumar
Sapna Langyan
Editors

Maize: Nutrition
Dynamics
and Novel Uses
Editors
Dharam Paul Chaudhary Sandeep Kumar
Directorate of Maize Research Germplasm Evaluation Division
New Delhi, India National Bureau of Plant Genetic Resources
New Delhi, India

Sapna Langyan
Directorate of Maize Research
New Delhi, India

ISBN 978-81-322-1622-3 ISBN 978-81-322-1623-0 (eBook)

DOI 10.1007/978-81-322-1623-0
Springer New Delhi Heidelberg New York Dordrecht London

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# Springer India 2014

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Foreword

Maize is a globally important cereal with the high production and

productivity. It is being cultivated in more than 160 countries under different
agro-climatic conditions. In India, it ranks third after wheat and rice. Maize
was primarily used as a food, but nowadays it is also used for feed and
industrial purposes. Therefore, apart from providing nutrients for humans
and animals, it also serves as a basic raw material for a number of industrial
products like starch, oil and protein, alcoholic beverages, food sweeteners
and biofuel. Because of its worldwide distribution and wider range of uses,
maize possesses many advantages over other cereals.
In India, maize occupies a prominent position, as each plant part finds
some specific application in feed, food or industrial sector. After harvest of
the grain, the dried stems are used as forage for ruminants. It gives the highest
average grain yield as compared to other cereals such as wheat and rice.
Nutritionally, maize contains about 6773 % starch, 713 % protein and
26 % oil. However, the protein quality of maize is poor because of the
deficiency of essential amino acids such as lysine and tryptophan. Quality
protein maize (QPM) is nutritionally improved as it contains required
concentrations of lysine and tryptophan. Among cereals and millets, oil is
extracted mainly from maize, and efforts have been made under National
Maize Improvement Programme towards the development of high-oil maize
hybrids with better oil quality. Maize oil is very popular and widely used
for human consumption as vegetable oil due to its cholesterol lowering
characteristics. The embryo which forms about 1012 % of the whole grain
is a rich source of oil.
Yellow maize is a rich source of vitamin A. Maize has more riboflavin
than wheat and rice and is rich in phosphorus and potash as well. Corn starch
is used as a thickening agent in the preparation of many edibles like soups,
sauces and custard powder. Corn syrup is used as an agent in confectionary
units. Corn sugar (dextrose) is used in pharmaceutical formulations, as a
sweetening agent in soft drinks, etc. Gel made from corn starch is used as
a bonding agent for ice cream cones due to its moisture retention
characteristics.

v
vi Foreword

It is well understood that the nutritional quality of maize is very important,

and there is still a need to improve it further. A sound strategy and well-
defined action plan is required to address these issues. The book entitled
Maize: Nutrition Dynamics and Novel Uses will provide valuable informa-
tion about the nutritional quality as well as researchable issues in the devel-
opment of nutritionally improved maize.

Dr. J.S. Sandhu

Agriculture Commissioner
Department of Agriculture & Cooperation
Ministry of Agriculture
New Delhi, India
Preface

Maize or corn (Zea mays) is a plant belonging to Poaceae, the family of

grasses. It is cultivated globally being one of the most important cereal crop
worldwide. Maize is not only an important human food but also a basic
element of animal feed and raw material for manufacturing a variety of
industrial products. It is a versatile crop grown over a range of agro-climatic
zones from 58 N to 40 S, from below sea level to altitudes higher than
3,000 m and in areas with 250 mm to more than 5,000 mm of rainfall per
year. Its growing cycle ranges from 3 to 13 months. The global maize
production during 2010 was 844 mt from an area of about 162 million
hectares. The USA, China, Brazil, Mexico, Argentina, Indonesia, India,
France, South Africa and Ukraine are the top ten maize-producing countries.
In India, maize is the third most important crop grown after rice and wheat.
Maize is consumed as a staple food in Africa, South America and some
parts of Asia. The nutritional quality of maize is poor as it is deficient in some
essential amino acids such as lysine and tryptophan. A nutritionally superior
maize named quality protein maize (QPM) has already been developed as a
result of nearly half a century of research dedicated to malnutrition eradica-
tion. Compared with traditional maize types, QPM has twice the amount of
lysine and tryptophan, as well as protein bioavailability that rivals milk
casein. QPM could be a practical food for alleviating protein malnutrition
in the developing countries, particularly sub-Saharan Africa and other parts
of the world that are worst affected by protein-energy malnutrition. Maize is
one of the most important natural multipliers of starch. Within just 3 months,
a single corn kernel produces more than 500 kernels containing about 70 %
starch. Maize is also a source of good-quality oil which is highly regarded for
human consumption as it contains essential fatty acid (linoleic acid) and
vitamin E which is an important antioxidant. Maize is also a rich source of
natural pigments, particularly carotenoids, which have diverse health benefits
ranging from maintaining normal vision to lowering of oxidative stress.
Maize is the most suitable crop for biofortification, particularly iron and
zinc, along with provitamin A carotenoids, for alleviating micronutrient
deficiencies.
Considering the nutritional role of maize worldwide in alleviating protein-
energy malnutrition (PEM), micronutrient deficiencies, vision impairments
and oxidative stress, it seems that maize is going to play a major role in
human nutrition apart from its utilization as an industrial raw material. Till
date, no comprehensive efforts have been made to discuss the nutritive

vii
viii Preface

value of maize. The present book provides a platform to understand the

various aspects of maize nutrition as a whole and its correlation with other
biotic and abiotic stresses. It also covers the role of natural variability
for various traits available in the germplasm and their future significance.
The value-added and fermented products of maize have also been discussed
thoroughly.
We pay our sincere thanks to Dr. O.P. Yadav, Director, Directorate of
Maize Research, for his valuable guidance and suggestions which helped us
in improving this book. We are also thankful to Dr. R. Sai Kumar and
Dr. Sain Dass, Ex-Directors, Directorate of Maize Research, for their prompt
help and motivation. Finally, we thank all the contributors for their pain-
staking efforts in the timely submission of their chapters.

New Delhi, India Dharam Paul Chaudhary

Sandeep Kumar
Sapna Langyan
Contents

Part I Introductory Chapter

1 Nutritive Value of Maize: Improvements, Applications

and Constraints . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
D.P. Chaudhary, Sandeep Kumar, and O.P. Yadav

Part II Protein Quality of Maize

2 Breeding Challenges and Perspectives in Developing

and Promoting Quality Protein Maize Germplasm . . . . . . 21
S.K. Vasal
3 Maize Protein Quality and Its Improvement:
Development of Quality Protein Maize in India . . . . . . . . . 37
M.L. Lodha
4 Molecular Interventions for Enhancing the Protein
Quality of Maize . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
Yogesh Vikal and J.S. Chawla

Part III Nutritional Quality of Maize

5 Maize: Grain Structure, Composition, Milling,

and Starch Characteristics . . . . . . . . . . . . . . . . . . . . . . . . . 65
Narpinder Singh, Amritpal Kaur, and Khetan Shevkani
6 Oil Improvement in Maize: Potential and Prospects . . . . . 77
Naveen Singh, Sujata Vasudev, D.K. Yadava,
D.P. Chaudhary, and K.V. Prabhu
7 Maize Carotenoid Composition and Biofortification
for Provitamin A Activity . . . . . . . . . . . . . . . . . . . . . . . . . . 83
Sandeep Kumar, Seema Sangwan, Rakesh Yadav,
Sapna Langyan, and Mohar Singh

ix
x Contents

Part IV Biotic and Abiotic Stresses in Maize

8 Insect-Pests and Their Management: Current Status

and Future Need of Research in Quality Maize . . . . . . . . . 95
M.K. Dhillon, V.K. Kalia, and G.T. Gujar
9 Physiological Response of Maize Under Rising
Atmospheric CO2 and Temperature . . . . . . . . . . . . . . . . . 105
Anjali Anand, Sangeeta Khetarpal, and Madan Pal Singh

Part V Value Addition in Maize

10 Maize Utilisation in Food Bioprocessing: An Overview . . . 119

Seema Sangwan, Sandeep Kumar, and Sneh Goyal
11 Maize Malting: Retrospect and Prospect . . . . . . . . . . . . . . 135
D.P. Chaudhary, D. Kumar, R.P.S. Verma,
Sapna Langyan, and Seema Sangwan
12 Value Addition in Maize . . . . . . . . . . . . . . . . . . . . . . . . . . 141
V.K. Yadav and P. Supriya
13 Fodder Quality of Maize: Its Preservation . . . . . . . . . . . . . 153
D.P. Chaudhary, S.L. Jat, R. Kumar, A. Kumar, and B. Kumar

About the Editors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161

 Part I
Introductory Chapter
 Nutritive Value of Maize:
Improvements, Applications 1
and Constraints

D.P. Chaudhary, Sandeep Kumar, and O.P. Yadav

Abstract
Maize is a globally important crop mainly utilized as feed, food and raw
material for diverse industrial applications. Among cereals, it occupies the
third place after wheat and rice and is a staple food for a large segment of
population worldwide, particularly in the Asian as well as African
countries. Its nutritional quality is, however, poor due to deficiency of
two essential amino acids, viz. tryptophan and lysine. The discovery of
opaque-2 gene has revolutionized the research in enhancing nutritional
quality of maize, and subsequent research efforts gave birth to the present-
day quality protein maize (QPM). This brings about a twofold increase in
the levels of lysine and tryptophan as the zein or prolamine fraction is
reduced by about 50 %. Starch is the major nutritional component of
maize kernel constituting about 70 % of its weight. Starch composition in
maize is genetically controlled, and significant variation has been
observed in the amylose to amylopectin ratio which makes it suitable
for different industrial purposes. Maize is also a source of oil which is
highly regarded for human consumption as it reduces the blood choles-
terol concentration. Many value-added products as well as fermented
foods have been produced from maize which is consumed in different
forms worldwide. Naturally, maize is a rich source of carotenoids such as
beta-carotene, zeaxanthin, lutein and cryptoxanthin which have highly
diverse health benefits ranging from maintaining normal vision to lower-
ing of oxidative stress. Efforts have been made towards the development
of biofortified maize rich in iron, zinc and provitamin A concentration.

1.1 Introduction

Maize (Zea mays L.), also known as corn, belongs

D.P. Chaudhary (*)  O.P. Yadav
Directorate of Maize Research, New Delhi, India
e-mail: chaudharydp@gmail.com
S. Kumar
Germplasm Evaluation Division, National Bureau
of Plant Genetic Resources, New Delhi, India
to the tribe Maydeae of Poaceae (Gramineae),
commonly known as grass family. It is a tall
monoecious plant domesticated by indigenous
peoples in Mesoamerica in the prehistoric times,
when grown in the form of a wild grass called

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 3
DOI 10.1007/978-81-322-1623-0_1, # Springer India 2014
4 D.P. Chaudhary et al.

a b
Yield (t/ha) Production (mt)
900
5.5 Maize Rice Wheat Maize Rice Wheat 844.40
850
5.2
5.0
800
4.5
4.8
4.4 750
713.70
4.0 700
4.1 672.02
3.5 634.40
650
650.88
3.0 3.0 600 626.87
2.5 2.9 550

2.0 500
2005 2006 2007 2008 2009 2010 2005 2006 2007 2008 2009 2010

Fig. 1.1 Maize productivity (a) and production (b) along with other major cereals in the world during 2010 (Source:
Based on FAO 2012)

teosinte (Beadle 1939). The history of modern-
day maize begins at the dawn of human agricul-
ture about 10,000 years ago. Over time, the sys-
tematic selection of certain varieties for their
desired traits led to the gradual transformation of
teosinte to its present-day form known as maize.
Today, maize is the worlds leading crop widely
cultivated as a cereal grain. It is cultivated in more
than 160 countries over a range of agroclimatic
zones. Being a C4 plant, maize has a specialized
physiology known as kranz anatomy, which
makes it well suited to hot and dry climates.
However, the extensive breeding interventions
made this plant suitable for colder regions as
well. It is grown from 58 N to 40 S, from below
sea level to altitudes higher than 3,000 m and in
areas with 250 mm to more than 5,000 mm of
rainfall per year (Dowswell et al. 1996). Its global
production is higher than that of rice and wheat
(Fig. 1.1). The global maize production during
2010 was 844 mt from an area of about 162
million hectares. The USA, China, Brazil,
Mexico, Argentina, Indonesia, India, France,
South Africa and Ukraine are the top ten maize-
producing countries (FAO 2012). The USA is the
largest producer and consumer of maize
contributing around 316 mt to the total global
maize production during 2010 followed by
China with a production of 177 mt (FAO 2012).
The maize production in Brazil and India was 56
and 21.72 mt, respectively, in 2010.
Globally, maize is used for feed followed by
food and as an industrial raw material. Maize is
the main staple food for people in Africa, South
America and some parts of Asia and is also used
worldwide as a fodder crop for livestock. Recent
advancements towards biofortification have led to
the development of quality protein maize (QPM)
and maize with higher provitamin A activity
which can play a major role in alleviating globally
the protein-energy malnourishment and vitamin A
deficiency, respectively.
This book covers various nutritional aspects of
maize including its importance and limitations as
food for humans, as livestock fodder, industrial
applications along with value addition, impor-
tance of its oil in human nutrition, various
fermented products and bioethanol production
and constraints in increasing its productivity in
the changing climatic scenery as discussed in the
following sections.
1.2 Composition of Maize Kernel
The maize plant is one of natures greatest
multiplier. It is a high-capacity factory for effi-
ciently converting large amounts of radiant energy
1 Nutritive Value of Maize: Improvements, Applications and Constraints
Fig. 1.2 Structure of
maize kernel
from the sun into stable chemical energy. This
energy is stored as cellulose, oil and starch in the
corn plant and its kernel. Approximately 4 months
after planting, a single kernel yields from 300 to
more than 800 kernels. Maize kernels develop
through accumulation of the products of photo-
synthesis, root absorption and plant metabolism
on the female inflorescence called the ear.
It consists of four major physical structures: endo-
sperm, germ or embryo, outer seed coat or peri-
carp and the tip cap (dead tissue found where the
kernel joins the cob) as shown in Fig. 1.2. The
pericarp is the outermost layer that is
characterized by high crude fibre content, mainly
consisting of hemicellulose, cellulose and lignin.
Endosperm is the largest component
(8085 %), followed by germ (910 %) and
pericarp (56 %). It is mainly composed of starch
(70 %) followed by small concentrations
(810 %) of protein (Lawton and Wilson 1987;
Prasanna et al. 2001). Fat content of the endo-
sperm is relatively low. The endosperm is com-
posed of a large number of cells, each packed
with starch granules embedded in a continuous
matrix of protein. The cell wall consists of non-
starchy polysaccharides (-glucan and
arabinoxylan), proteins and phenolic acids. Pro-
tein bodies are composed almost entirely of a
prolamine-rich protein fraction known as zein.
Maize grain has two types of endosperm floury
and horny. Floury endosperm contains loosely
packed starch granules surrounding the central
5
fissure, while horny endosperm has tightly
packed, smaller starch granules towards periph-
ery. The germ contains high concentrations of fat
(33 %) and relatively high levels of protein
(1819 %) and minerals (Watson 1987). Germ
oil is relatively stable due to the presence of
natural antioxidants and highly regarded for
human consumption because of its fatty acid
composition, which mainly consists of oleic and
linoleic acids. Maize contains various bioactive
constituents, such as carotenoids, anthocyanins,
tocopherols and phenolic compounds that have
many health-promoting and disease-preventing
properties.
1.3 Nutritional Quality of Maize
Improving nutritional quality in cereal crops is
particularly important as the benefits can easily
spread to hundreds of millions of people in a
most rapid and effective manner without chang-
ing the traditional food habits. A significant
human population consumes maize as a staple
food, worldwide, the nutritional quality of
which in turn depends upon the chemical com-
position of various components of its kernel.
Since endosperm accounts for the largest compo-
nent, the quality of maize kernel as a whole,
therefore, depends largely upon its chemical
composition.
6 D.P. Chaudhary et al.
1.3.1 Protein Quality
Protein quality, from a nutrition perspective, is a
comparative term used to describe how well a
food protein fulfils the bodys requirements and,
therefore, how useful the protein is for the
biological system. This is determined by the
building blocks which make up the protein, called
amino acids. Among the 20 primary amino acids,
nine are considered essential as these cannot be
synthesized by our body and so must come from
the diet. The best quality protein is one which
provides essential amino acid pattern very close
to that of the tissue proteins. Usually, milk and egg
proteins serve as reference protein, because of
their superior quality. As per Joint FAO/WHO
Expert Consultation (1991) recommendations,
the essential amino acid composition (mg/g pro-
tein) of reference protein required for 25-year-
old children is as follows: lysine 58, threonine 34,
tryptophan 11, methionine + cysteine 25. Various
methods have been introduced to measure protein
quality including biological value (BV), net pro-
tein utilization (NPU) and protein efficiency ratio
(PER). The maize protein quality is, however,
poor due to deficiencies of two main essential
amino acids, lysine and tryptophan, and excess
of leucine in the endosperm protein of its kernel
(Osborne and Mendel 1914). The endosperm pro-
tein is classified into various fractions such as
albumin (3 %), globulin (3 %), zein/prolamine
(60 %) and glutelin (34 %). Based on their solu-
bility, genetic properties and the apparent molec-
ular masses, zeins have been classified into - (22
and 19 kDa), the most abundant - (14 kDa), -
(27 and 16 kDa) and -zein (10 kDa) (Wilson
et al. 1981). The -zein fraction is rich in cysteine
while and fractions are rich in methionine.
Generally in normal maize, zein fraction contains
higher proportion of leucine (18.7 %), phenylala-
nine (5.2 %), isoleucine (3.8 %), valine (3.6 %)
and tyrosine (3.5 %), but low amounts of other
essential amino acids such as threonine (3 %),
histidine and cysteine (1 %), methionine (0.9 %)
and lysine (0.1 %), but is devoid of tryptophan.
The non-zein protein fraction is balanced and rich
in lysine and tryptophan. The high proportion of
zeins in the endosperm is the primary reason for
the poor protein quality of maize protein (Vasal
2000).
The discovery of opaque-2 gene and its associ-
ation with higher lysine and tryptophan content in
the maize endosperm led to the beginning of the
development of QPM (Mertz et al. 1964). High-
lysine mutants such as opaque-2 were developed
and introduced into normal maize which brings
about a twofold increase in the lysine and trypto-
phan levels. Further, lysine-deficient zein or prola-
mine fraction is reduced dramatically by about
50 %, while other fractions, such as albumins,
globulins and glutelins which are rich in lysine,
show a marked increase. The soft endosperm
opaque-2 maize was, thus, evolved. However, the
opaque-2 maize varieties could not become popu-
lar due to its soft and chalky grains texture, inferi-
ority in terms of yield and agronomic performance
and susceptibility to insects and pest infestation.
This was the big setback for researchers involved
in the development of nutritionally improved
maize. However, later on, some partially hard
endosperm or modified grains had been
observed which possess desired concentrations of
essential amino acids. The modified opaque-
2 maize with hard endosperm is known as QPM.
1.3.2 Carbohydrate Profile
Major carbohydrate of maize kernel is starch,
which contributes approximately 70 % of the ker-
nel weight. In fact, maize is one of the greatest
manufacturers of starch. The starch molecule is a
homopolymer of repeating anhydroglucose units
joined by -glycosidic linkages, the aldehyde
group of one unit being chemically bound to a
hydroxyl group on the next unit through hemiace-
tal linkages. The 1,4-linkages yield straight-chain
starch molecules called amylose, while the 1,6-
linkages serve as the branching point in branched-
chain starch molecules called amylopectin. The
starch composition in maize is genetically con-
trolled. In normal maize, amylose makes up
2530 % of the starch and amylopectin up to
7075 %. Wide genetic variability exists in
1 Nutritive Value of Maize: Improvements, Applications and Constraints
maize with respect to starch profile. On the basis
of starch composition, maize is categorized in
three classes: (1) waxy maize, which contains
almost 100 % amylopectin starch; (2) high-
amylose maize, with starch containing amylose
content between 40 % and 70 %; and (3) sugary
maize, containing lower starch but higher level
of sucrose (Nelson and Pan 1995). Maize with
amylose as high as 85 % (amylomaize) is also
reported. These variants have different industrial
applications. Waxy maize is having large stake in
food industry, whereas high-amylose maize is
preferably required by textile industry. Waxy
mutants starch has been reported to be more crys-
talline than regular cereal starches (Singh et al.
2003; Vandeputte et al. 2003). Dietary starch
varies greatly in digestibility and its effects on
the utilization of other nutrients. Maize possesses
wide genetic variability with respect to digestibil-
ity of its starch molecule. Starch is classified into
three groups depending upon the rate of release
and absorption of glucose in the gastrointestinal
tract: rapidly digestible starch (RDS), slowly
digestible starch (SDS) and resistant starch (RS).
RDS is the group of starches that can be rapidly
hydrolyzed by digestive enzymes; SDS is the
group that is digested at a relatively slow rate
(Englyst et al. 1992). However, a part of it is not
digested in the small intestine and reaches the
large intestine (colon) where it is fermented by
the gut microflora that produces short-chain fatty
acids as end products which are known to promote
the optimal function of the viscera (Topping and
Clifton 2001). This indigestible portion of starch
is known as resistant starch. Waxy maize starch is
more rapidly digested than high-amylose starch,
attributed to more surface area per molecule of the
amylopectin than amylose. Many health benefits
such as improved cholesterol metabolism and
reduced risk of type II diabetes and colon cancer
have been associated with the consumption of RS
(Hoebler et al. 1999).
Major food uses for starch include sweeteners,
brewing adjuncts, chemicals/pharmaceuticals,
viscosity control agents in canning, starch-based
confectionery products and bakery applications.
Non-food industrial applications include
7
adhesives in paper and building materials and
coatings and sizings in textiles and paper products
(Kulp 2000). The application of starch in various
industries is primarily determined by its functional
properties such as viscosity, swelling, solubility,
gelatinization, pasting and retrogradation, which
vary considerably from crop to crop and with
ecological and agronomic influences (Riley et al.
2006; Yuan et al. 2007). Raphael et al. (2011)
compared the functional properties of two major
botanical sources of starches, maize and cassava
for East African starch industries, and found
applications in different industries. Most of the
corn starch is extracted from the kernel through
wet milling process along with other by-products.
Subsequently, the starch fraction can be processed
through chemical or biochemical procedures to
enhance its applicability in food and industrial
products. For example, oxidized starches are
used in laundry starches and paper manufacturing.
Dextrins (dry heating and roasting of starch with
or without an acid or alkaline catalyst and process
known as dextrinization) are widely used as
quick-setting pastes in paper products. Instantized
products such as instant puddings are produced
from pregelatinization of starches. Starches are
treated with enzymes to produce high-fructose
corn sweeteners found in soft drinks. It can also
be fermented to produce alcohol. Starch can be
derivatized by a number of agents reacting with
hydroxyl groups, cross-linking and stabilization
derivatives and other derivatives depending upon
required functional properties such as increased
water-combining capacity, impeded retrograda-
tion, etc. (Jackson 1992; Kulp 2000). Further, as
starch is totally biodegradable, it can be used for
developing bioplastics in the form of packaging
material, fast-food service ware, etc.
Maize kernel also possesses some complex
carbohydrates. The complex carbohydrate con-
tent of the maize kernel comes from the pericarp,
tip cap, endosperm cell walls and to a smaller
extent the germ cell walls. Maize bran is com-
posed of 75 % hemicellulose, 25 % cellulose and
0.1 % lignin on a dry-weight basis (Sandstead
et al. 1978; Van Soest et al. 1979). These
constituents, though indigestible, help in
8 D.P. Chaudhary et al.
normalizing the digestive process. Other
carbohydrates are simple sugars present as glu-
cose, sucrose and fructose in amounts ranging
from 1 % to 5 % of the kernel weight. Immature
kernels contain relatively high levels of sugars
and fewer amounts of other nutritional com-
ponents which accumulate during development
(Boyer and Shannon 1982). The sugar content
makes the most clearly recognizable component
of sweet maize quality as sweet corn is eaten at
an immature stage of development (Evensen and
Boyer 1986).
1.3.3 Oil Composition
Oil is mainly confined to the germ which provides
around 85 % of the total kernel oil. The rest of the
oil is dispersed in endosperm and hull fractions.
Maize oil is mainly used in cooking and has high
smoke point which makes it valuable for frying
purposes (Katragadda et al. 2010). It is also a key
ingredient in some margarines. Oil is an important
by-product of starch industry. Normal maize
provides around 26 % of oil, whereas high-oil
maize contains more than 6 % of oil (Lambert
2001). High-oil maize differs in kernel composi-
tion as its germ size is found to be larger as com-
pared to normal maize. Since most of the oil is
contributed by the germ, therefore, increasing oil
content in maize is directly proportional to the
germ size (Motto et al. 2005). Increased germ
size, however, is associated with reduction in
starch as both oil and starch are negatively
correlated (Yang et al. 2013). Maize genotypes
with higher oil concentrations are of interest to
many livestock feeders, primarily because of the
higher calorific value of the grain (Lambert 2001).
High-oil varieties of maize were developed at the
University of Illinois (Dudley and Lambert 1992).
Although these genotypes possess higher
energy content, they showed poor agronomic
characteristics.
Maize oil is valued because of its low levels of
saturated fatty acid, i.e. on an average 11 %
palmitic acid and 2 % stearic acid. On the other
hand, it contains high levels of polyunsaturated
fatty acids (PUFA), mainly linoleic acid
(24 %), and small concentrations of linolenic
acid. Corn oil plays an important role in human
nutrition. The PUFAs help in regulating blood
cholesterol and lowering the elevated blood pres-
sure (Hauman 1985; Dupont et al. 1990). Corn
oil is a rich source of linoleic acid which is an
essential fatty acid that the body cannot synthe-
size. The term essential fatty acid refers to the
fatty acids, required for biological processes.
Only two fatty acids are known to be essential
for humans: alpha-linolenic acid (an omega-3
fatty acid) and linoleic acid (an omega-6 fatty
acid). The Food and Agriculture Organization
and World Health Organization recommend
about 24 % of energy in the form of essential
fatty acids, with an additional 3 % energy for
pregnant or breast-feeding women. A tablespoon
serving of corn oil is sufficient to satisfy the daily
essential fatty acid requirement of a healthy child
or an adult. Corn oil is also recognized as an
excellent source of tocopherols. Tocopherols
function as antioxidants and provide a good
source of vitamin E. Like essential fatty acids,
vitamin E also represents an essential component
of human diet as it cannot be synthesized in the
body. It is a class of strong antioxidants which
protect polyunsaturated fatty acids in membranes
against degradation by reactive oxygen species
such as ozone, singlet oxygen, peroxides and
hyperoxides (Dormann 2003). So, the antioxi-
dant activity of tocopherols is important not
only from health point of view but also in terms
of oil quality as it helps in increasing its shelf life
by retarding the development of rancidity. The
four major tocopherols found in corn oil are
alpha-, beta-, gamma- and delta-tocopherol. In
commercially available corn oil, gamma-
tocopherol is the most abundant, followed by
alpha-tocopherol and delta-tocopherol. The
tocopherol that exhibits the greatest antioxidant
effect is delta-tocopherol, whereas alpha-toc-
opherol has the highest vitamin E activity.
1.4 Natural Pigments
The colour of maize ranges from white to
yellow, red, blue, purple, etc. Blue-, purple- and
red-pigmented maize kernels are rich in
anthocyanins with well-established antioxidant
1 Nutritive Value of Maize: Improvements, Applications and Constraints
and bioactive properties (Adom and Liu 2002).
Anthocyanin, carotenoid and phenolic contents in
maize vary with the colour. The highest concen-
tration of anthocyanin pigments in maize is present
in the pericarp portion, whereas the aleurone layer
contains small amounts (Moreno et al. 2005).
Maize exhibits considerable natural variation for
kernel carotenoids with some genotypes
accumulating as high as 66.0 g/g (Harjes et al.
2008). The predominant carotenoids in maize
kernels in decreasing order of concentration are
lutein, zeaxanthin, -carotene, -cryptoxanthin
and -carotene. Yellow maize has more caro-
tenoids than floury maize. Generally, provitamin
A carotenoids constitutes only 1020 % of total
carotenoids in maize whereas 3050 % represent
by zeaxanthin and lutein each. In typical maize,
concentrations of provitamin A carotenoids, i.e.
-carotene, -carotene and -cryptoxanthin, range
from 0 to 1.3, 0.13 to 2.7 and 0.13 to 1.9 nmol/g,
respectively (Kurilich and Juvik 1999).
Maize is also known to contain a wide range
of phenolic acids. Ferulic acid is an important
phytochemical in maize and its concentration
varies in different maize types. The high-carotenoid
maize contains higher amount of total ferulic acid
compared to white, yellow, red and blue maize.
Most of the ferulic acid in maize is present in
bound form (Adom and Liu 2002). The bulk of
phenolics (phenolic acids, flavonoids and conju-
gated amines) are concentrated in the pericarp and
aleurone layers as well as the germ, with traces in
the endosperm (Sen et al. 1994).
1.5 Mineral Composition
Iron deficiency is the most common and wide-
spread nutritional disorder in the world. Apart
from affecting a large number of children and
women in developing countries, it is the only
nutrient deficiency which is also significantly
prevalent in industrialized countries. The num-
bers are staggering: two billion people over
30 % of the worlds population are anaemic,
many due to iron deficiency, and in resource-
poor areas, this is frequently exacerbated
by infectious diseases. It can retard mental devel-
opment and learning capacity and impair
9
physical growth during childhood and adoles-
cence, while in adults it reduces the capacity to
do physical labour (Bouis 2002). Deficiency of
zinc, another common micronutrient, lowers the
intestinal absorption of fat and fat-soluble
vitamins including retinol (Ahn and Koo 1995a,
b; Kim et al. 1998). Similarly, other mineral
elements, including calcium, copper, magne-
sium, manganese, phosphorus and potassium,
are also considered essential for various physio-
logical activities including growth and develop-
ment of bones, teeth, blood, nerves and skin;
synthesis of vitamins, enzymes and hormones;
as well as for healthy functioning of the nervous
system, blood circulation, fluid regulation, cellu-
lar integrity, energy production and muscle con-
traction (MacDowel 2003; ODell and Sunde
1997).
Mineral composition of cereal grains can be
affected by a number of factors such as soil type
and fertility, soil moisture, environmental
factors, crop genotype and interactions among
nutrients (Arnold et al. 1977; Feila et al. 2005).
Wide natural variability for various minerals
exists in the maize germplasm. Menkir (2008)
evaluated a vast set of tropical-adapted inbred
lines for mineral concentrations. The best inbred
lines identified from each trial had 3278 % more
Fe and 14180 % more Zn than their trial aver-
age. The first two principal component axes,
which accounted for 5564 % of the total varia-
tion in kernel mineral concentrations, stratified
the inbred lines in each trial into four groups
based on differences in their grain mineral
compositions. None of the correlations of Fe
and Zn with Mn, Cu, Ca, Mg, K, P and S were
significant and negative in the various trials,
while the correlations of Fe with Zn were posi-
tive and significant. Ullah et al. (2010) also
analysed grains of ten maize varieties for mineral
composition and reported the range of Na
(540.30620.41 ppm), K (29153471 ppm), Ca
(410590 ppm), Fe (38.0256.14 ppm), Zn
(37.0552.4 ppm), Mg (985.21125.3 ppm) and
Cu (11.0214.25 ppm). High levels of phytate
(myo-inositol hexaphosphate) are present in
unrefined cereals, which is a powerful inhibitor
of iron and zinc absorption in both adults
(Egli et al. 2004; Mendoza et al. 1998) and
10
children (Davidsson et al. 2004). Any reduction
in dietary phytate can have a positive effect on
zinc absorption (Lonnerdal 2002) and iron
absorption (Mendoza et al. 1998).
1.6 Biofortification of Maize
Globally, approximately one-third of preschool-
age children and 15 % of pregnant women are
estimated to be vitamin A deficient (WHO
2009). The problem becomes more severe partic-
ularly in the developing countries whose poor
populations rely on a single staple crop for their
sustenance, e.g. Africa and Southeast Asia have
the highest burden of vitamin A deficiency (WHO
2009). The consequences of vitamin A deficiency
include blindness, reduced growth in children and
increased morbidity and mortality (Sommer and
West 1996; Shankar et al. 1999; Rice et al. 2004;
Maida et al. 2008). Further, interaction of vitamin
A content with both iron as well as zinc content
has been well documented (Hess et al. 2005). Iron
metabolism is negatively affected by vitamin A
deficiency in the diet, and iron is not incorporated
effectively into haemoglobin (Hodges et al. 1978).
The co-occurrence of iron and vitamin A
deficiencies has been found in infants in South
Africa (Oelofse et al. 2002), preschool children
in the Marshall Islands (Palafox et al. 2003),
school-age children in Cote dIvoire (Hess 2003)
and pregnant women in India, Nepal and Malawi
(Pathak et al. 2003; Dreyfuss et al. 2000; van den
Broek and Letsky 2000). Similarly, zinc intake
has been considered to be inadequate for an
estimated 30 % of the populations in 46 African
countries (Hotz and Brown 2004). Zinc defi-
ciency, as already, discussed lowers the intestinal
absorption of fat and fat-soluble vitamins includ-
ing vitamin A retinol in rats. Various implications
associated with iron and zinc deficiencies have
already been discussed along with other minerals
under mineral composition part.
Biofortification is the development of micronu-
trient-dense staple crops using the best traditional
breeding practices and modern biotechnology.
To alleviate the vitamin A and micronutrient,
D.P. Chaudhary et al.
particularly iron and zinc deficiencies from rural
areas of developing countries, biofortification of
major staple food crops of respective areas or
countries is the only feasible way, since this will
better ensure targeting and compliance. As the
third most important cereal staple food crop world-
wide (FAPRI 2009) and a major cereal staple for
African consumers (FAOSTAT 2010), maize
qualifies as a suitable crop for biofortification.
Before devising a strategy to enhance the provita-
min A carotenoids and iron and zinc micro-
nutrients in maize, one question that must be
addressed is the target quantity for biofortification.
This is related to the bioavailability or the fraction
of an ingested nutrient that becomes available to
the body for utilization in physiological functions
or for storage (Jackson 1997; Fraser and Bramley
2004). There are numerous factors that influence
bioavailability. After considering all these factors,
it has been decided that at least 15 g -carotene/g
dry kernel weight, 60 mg/kg iron and 55 mg/kg
zinc are required for biofortified maize to have an
impact on nutrition (Graham et al. 1999).
1.7 Value Addition
A broad definition of value addition is to
economically add value to a product and form
characteristics more preferred in the market
place. Value addition in maize has a great poten-
tial. There are several value-added products of
maize, particularly QPM and baby corn, which
not only increase the farm income but also pro-
vide employment to rural youth and farm women.
QPM has been used for the development of
traditional products, baked products, extruded
products, convenience foods, infant food, health
foods, snacks and savoury items, specialty foods,
etc. which can meet the nutritional need of vulner-
able section. Further, maize was blended with soya
bean/green gram in ratio 70:30 and the product
developed includes cake, biscuit, halwa, upma,
vada, etc. (Kawatra and Sehgal 2007). A number
of value-added products have been developed
from specialty corns taking in care the various
age groups and their requirements as discussed in
detail in related chapter of value addition.
1 Nutritive Value of Maize: Improvements, Applications and Constraints
1.8 Fermented Maize Foods
Fermented foods are considered as palatable and
wholesome foods prepared from raw or heated
raw materials and appreciated for their attributes
such as pleasant flavour, aroma, texture,
improved processing properties and better
digestibility. Maize is a good source of dietary
fibre and protein while being very low in fat and
sodium. The endosperm consists of approxi-
mately 70 % starch embedded in a protein matrix
that makes the maize an excellent substrate for
fermentation. Maize is processed, fermented and
consumed in various ways. Some of the
fermented maize products include Chicha (a
clear, yellowish, effervescent, alcoholic bever-
age having alcohol content between 2 % and
12 %, most popular in South and Central Amer-
ica), Tesguino (a slurry-like alcoholic beverage
prepared from germinated maize or maize stalk
juice commonly consumed in Northern and
Northwestern Mexico), Umqombothi (a pink,
opaque, mild alcoholic drink having yoghurt-
like flavour with thin consistency, prepared
from maize and sorghum popular in South Afri-
can population), Busaa (a Kenyan opaque maize
beer having 24 % ethanol and 0.51 % lactic
acid), Atole (a sour porridge-type product of
Southern Mexico), Pozol (fermented maize
dough formed into balls of various shapes and
sizes which is an important recipe of Southeast-
ern states of Mexico), Ogi (prepared from
submerged fermentation of maize or sorghum
and millet is a traditional weaning food common
in West Africa), Mahewu (a non-alcoholic sour
beverage made from corn meal and is mostly
consumed in Africa and some Gulf countries),
Pito (a traditional beverage, very popularly con-
sumed throughout Nigeria and Ghana) and Uji (a
sour maize gruel from East Africa).
1.9 Industrial Aspects
Maize is an important raw material for the
production of a number of industrial products
such as starch, oil, ethanol, malt and animal
11
feed. Two types of industrial processes are
common for maize processing: wet milling and
dry grinding. Wet milling involves separation of
grain kernels to its constituent components such as
germ, bran (fibre), protein and starch. In this pro-
cess, the grains are steeped or soaked into water
for 2448 h to facilitate the separation of kernel
components. These components are then sequen-
tially removed through a series of industrial pro-
cesses to obtain the final products. The germ is
removed and processed for the extraction of oil
which is then purified and sold for human con-
sumption. Bran (fibre components) is used in
the animal feed. The starch as well as gluten is
separated through centrifugation processes. Maize
is an important industrial raw material for the
production of starch which is utilized in a number
of industries such as paper industry, textile indus-
try, pharmaceutical industry, food industry and
leather industry. It can be processed further to
synthesize a number of components such as dex-
trose, fructose and high-fructose corn syrups.
Corn gluten meal is a by-product which is sold
to poultry sector as a high-protein feed.
In the dry grind process, the entire grain is
ground into flour (Butzen and Haefele 2008).
Dry grind process is usually employed for the
production of ethanol from maize. In this process,
kernels are ground and mixed with water to form
slurry called mash which is further cooked dur-
ing which the starch is enzymatically converted to
sugars. Sugars are fermented to ethanol by the
addition of suitable yeast. The ethanol is extracted
and the solid remains are dried to produce
distillers dried grain soluble (DDGS) which is
used as a high-protein ingredient in animal diet.
A significant part of maize produced is utilized for
the production of biofuels in the USA. In India,
policy makers are seriously considering the explo-
ration of maize for the synthesis of biofuels to
meet the ever-increasing demand of fuels for
vehicular traffic. The increasing production of
maize throughout the world has necessitated
finding economically viable industrial processes
for its conversion to ethanol for its use as a bio-
fuel. Another important use of maize could be its
conversion to malt to be used for the production of
beer. Presently, maize is used as an adjunct along
12
with barley which is the most preferred cereal for
malting purposes. The issue has been addressed in
a separate chapter.
1.10 Specialty Maize
Plant breeders have altered through genetics the
starch, protein and oil content of corn to meet the
needs of the human population, livestock feeder,
the food industry and other industrial users of
corn in a better way. As a result of their
modifications of ordinary dent types, new spe-
cialty corns have been created, including sweet
corn, waxy, high-amylose, high-oil and baby
corn. Sweet corn and baby corn are two impor-
tant special types which are gaining wide-scale
popularity in the recent times. Sweet corn is a
variety of maize with high sugar content. Unlike
other normal maize, which are harvested when
the kernels are dry and mature, sweet corn is
harvested when the kernels are immature (milk
stage) and eaten as a vegetable. Immature stan-
dard sweet corn is considered desirable when
kernels are succulent because of a mutant reces-
sive sugary-1 gene (su-1) that retards the conver-
sion of sugar into starch during endosperm
development. In comparison, sucrose produced
in leaves from photosynthesis in dent corn is
passed on to developing kernels, where it is rap-
idly converted to dextrin (a non-sweet, water-
soluble polysaccharide) and then to starch. The
sugary-1 gene slows down this process.
Baby corn is the young ear, harvested when
the silks have just emerged and no fertilization
has taken place. Baby corn is a very delicious and
nutritious vegetable and has been considered to
be a high-value agriculture produce in national
and international markets. It is free from the
harmful effects of pesticides as baby corn is
covered tightly with husk and due to early
harvesting; it is devoid of harmful pests and
diseases. A number of products with additional
value from baby corn have been developed, and
there is a vast scope of developing and promoting
value-added products from baby corn.
D.P. Chaudhary et al.
1.11 Maize as Fodder for Livestock
Green fodders are the most important component
of animal husbandry as they provide cheap and
nutritious feed for livestock. The economics of
milk production depends primarily upon the
availability of nutritious green fodders. Maize is
rated as one of the best non-legume fodder. It is
considered ideal forage because it grows quickly,
produces high biomass, is palatable, is rich in
nutrients and helps to increase body weight and
milk quality in cattle (Sattar et al. 1994). As
fodder for livestock, maize is an excellent, highly
nutritive and sustainable source (Hukkeri et al.
1977; Iqbal et al. 2006). Moreover, maize is
grown round the year and is also free from anti-
quality components. It contains high concen-
trations of protein and minerals and possesses
high digestibility (Gupta et al. 2004). Specialty
corn such as sweet corn and baby corn serves
dual purpose as they provide fodder along with
special purpose maize. Maize also possesses
excellent ensiling characteristics as it contains
sufficient quantities of soluble sugars required
for proper fermentation (Allen et al. 2003). In a
recent study carried out at Directorate of Maize
Research, excellent silage was produced from
baby corn stalks (after the harvest of baby corn)
after 45 days of ensiling (unpublished data). The
nutritional quality of maize as compared to other
forages, the silage-making technology and the
potential of specialty corn as fodder for livestock
are discussed in detail in a separate chapter.
1.12 Maize Under Changing
Climatic Scenario
Rise in atmospheric CO2 with parallel increase in
temperature is posing to be a serious threat to the
farming community. During the last 12 years, the
increasing rate of CO2 has been 1.9 l l 1 year 1
and is predicted to reach the level of 570 l l 1 by
the middle of this century (IPCC 2007). All the
physiological processes are affected by the
change in the environment that finally limits
1 Nutritive Value of Maize: Improvements, Applications and Constraints
productivity of the crop. Elevated CO2 causes
partial stomatal closure thereby decreasing leaf
transpiration, while at the same time, the carbon
assimilation is increased (Morison 1998). C4
plants possess two types of cells, mesophyll and
bundle sheath cells, and possess a mechanism
that concentrates CO2 in the bundle sheath cells
to levels that have been estimated to be 38 times
more than the atmospheric CO2 concentration
(Kanai and Edwards 1999). A differential
response to increased CO2 levels was observed
in maize with studies showing no enhancement
in growth (Hunt et al. 1991) to 50 % stimulation
(Rogers and Dahlman 1993). High temperature
denatures RuBisCO activase rendering it unable
to fit correctly onto RuBisCO. Consequently, inac-
tive RuBisCO is not converted to active form
(Crafts-Brandner and Salvucci 2000). Further,
high temperature reduces grain size due to steady
decrease in the duration of grain filling combined
with a failure of compensation by increased rate
of dry matter accumulation above a threshold
temperature. In respective chapter, various
components affecting the growth and yield of
maize and other cereal crops under elevated CO2
and temperature are discussed in detail.
1.13 InsectPest Scenario
The grain yields of traditional maize genotypes in
India are quite low, and in spite of sincere efforts,
the yield potential of Indian maize genotypes has
not reached even half the mark to that in the USA.
Furthermore, the gap in yield potential of maize
genotypes under experimental conditions and that
under farmers fields is huge, due to different
stress factors; mainly, the insect pests during veg-
etative stage need greater attention to break this
yield gap. Different insect pests, viz. maize stalk
borer, pink stem borer, sugarcane leafhopper,
shoot bug, armyworm, shoot fly, corn leaf aphid,
cob borer and termites, have constrained the
increase in yield potential of the maize genotypes
deployed in India. It is likely that with the increase
in concentration and quality of nutritional
compounds, particularly the protein quality in
QPM, maize might favour the proliferation of
13
insect pests as they too prefer quality food for
their growth and development and could be a
major constraint to increasing production and pro-
ductivity of QPM. The maize stalk borer, Chilo
partellus, alone causes 26.780.4 % yield loss
under different agroclimatic conditions in India
(Reddy and Zehr 2004). Host plant resistance is
one of the effective means of minimizing losses
due to insect pests. However, most of the maize
varieties and hybrids released for cultivation are
potentially susceptible to C. partellus during veg-
etative stage (Kumar 1997) and maize weevils,
Sitophilus spp., under field and storage conditions
(Arnason et al. 1993; Hossain et al. 2007). Dam-
age potential of different insect pests, status of
host plant resistance and the mechanisms of resis-
tance involved and the management of major
insect pests have also been discussed in the related
chapter.
1.14 Disease Scenario
Maize crop is affected by a total of 62 diseases
worldwide. Out of these 62, 16 have been
identified as a major constraint in increasing pro-
duction and productivity of maize in India.
Banded leaf and sheeth blight (BSLB), pythium
stalk rot, bacterial stalk rot, post-flowering stock
rot (PFSR), polysora rust and downy mildews are
the major threat to the potential yield of maize.
Global losses in maize due to diseases were
estimated about 9 % in 20012003 (Oerke
2005). These losses varied significantly by region
with estimates of 4 % in northern Europe and
14 % in West Africa and South Asia (http://
www.cabicompendium.org/cpc/economic.asp).
Losses have tended to be effectively controlled in
high-intensity agricultural systems where it has
been economical to invest in resistant germplasm
and pesticide applications. However, in areas like
Southeast Asia, hot, humid conditions have
favoured disease development, while economic
constraints prevent the deployment of effective
protective measures. Since maize is affected by a
number of diseases with different pathogenic
nature, no single management strategy is suffi-
cient to manage such complex group of diseases.
14
Damage potential of different diseases, status of
host plant resistance and the mechanisms of
resistance involved, and the management of
major disease are needed to be addressed by an
integrated disease management approach.
1.15 Future Strategies
for Nutritionally Enriched
Maize
Maize industry is growing and will attain more
importance in the future due to its diverse
applications. Maize could serve as an important
food source for the increasing human population
worldwide. However, research interventions are
still needed to improve the nutritional quality and
to popularize the nutritionally improved maize.
The nutritional superiority of QPM over normal
maize is well established. QPM can become a
boon to vulnerable groups particularly from rural
poor populations worldwide. QPM rather tastes
sweet compared to normal maize and, therefore,
can be consumed without any change in the tra-
ditional food habits. Recently, the Indian Gov-
ernment has set aside a significant amount in its
annual budget towards the development of nutri-
farms in the country ensuring that the farmers
growing QPM should get remunerative price for
their produce. There is a need to educate the
masses that QPM is a product of conventional
breeding and does not involve any transgene. A
lot of value-added products from QPM such as
QPM Mix-I, QPM Mix-II, QPM ladoo, honey
maize chocolate, maize coconut chocolate,
maize coconut toffee, maize groundnut toffee
and choco maize bar have been developed, and
efforts are needed to popularize these products.
The vulnerable groups, i.e. infants, preschool
children, pregnant and lactating mothers and
elderly people, are going to get the maximum
benefit from QPM and its products. Efforts
should be made towards the development of
maize germplasm containing high methionine
which is an important ingredient in the poultry
feed. Moreover, efforts should be made towards
understanding the mechanism of action of modi-
fier genes in the QPM kernel, the expression of
D.P. Chaudhary et al.
which is affected by genetic and environmental
factors.
Biofortification technology is highly sustain-
able and cost-effective as the once developed
fortified seeds can be grown time and again and
could easily reach the undernourished populations
in relatively remote rural areas, delivering natu-
rally fortified foods to people with limited access
to commercially marketed fortified foods that are
more readily available in urban areas. Maize
biofortification for provitamin A carotenoids
benefits human health and also adds to commer-
cial value of food as these are natural colourants.
National germplasm collections hold untapped
potential for maize improvement. Biofortification
of maize requires large-scale germplasm screen-
ing and utilization of identified trait-specific mate-
rial into the breeding programme. Further, the
developed biofortified maize material using trait-
specific germplasm should not only contain higher
quantities of provitamin A carotenoids and
minerals but also have all other crucial traits
including higher yield, insect pest and disease
resistance and better nutritional quality. So, there
is a need to tag new genes/loci for various traits.
Further, mutational studies can also provide an
insight about the regulatory points in the meta-
bolic pathways. Similarly, stage-specific metabo-
lite profiling and its correlation with candidate
gene expression could provide important informa-
tion regarding regulation chemistry and expres-
sion patterns. So, an interdisciplinary approach
including biochemistry, genetics, plant breeding
and nutrition is required for tagging of new genes
and identification of rate-limiting steps of the
pathways along with gene expression patterns
with respect to time and allozymic diversity,
which in turn might provide important informa-
tion for deciding futuristic strategies regarding
overall maize improvement.
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 Part II
Protein Quality of Maize
 Breeding Challenges and Perspectives
in Developing and Promoting Quality 2
Protein Maize Germplasm

S.K. Vasal

Abstract
The chapter discusses the development of nutritionally enhanced maize
popularly known as quality protein maize (QPM). The focus will be on
describing ever-evolving breeding options and strategies in developing
this maize. Most of the examples will be quoted from CIMMYTs work
where much of the research and practical breeding has been attempted.
Initially, approaches of CIMMYT breeders were not very different from
other researchers. High-lysine mutants such as opaque-2, floury-2,
opaque-7, and a few double mutant combinations of different endosperm
mutants were tried. Experiences, difficulties, and manipulation of differ-
ent mutants will be presented having relevance to the development of
high-lysine maize genotypes. The strengths and weaknesses of early
breeding efforts will be critically discussed revealing complexity of inter-
related problems affecting acceptance at producer, consumer, and indus-
trial processing level. Initial research efforts at CIMMYT and elsewhere
were confined to developing soft endosperm opaque versions of counter-
part normal maize varieties and hybrids. Extensive testing of such
materials exposed a series of problems of poor agronomic performance,
unacceptable kernel appearance, and some problems of specific nature
under certain environmental conditions. High hopes and early optimism of
mid-1960s and early 1970s received a major setback in research efforts
and brought lots of frustration and declining interest in high-lysine maize
types. Funding declined resulting in reduced breeding efforts and
abandoning of research partially or completely in several parts of the
world. Only a handful of institutions and breeding programs continued
research in a persistent and a systematic manner. Continuing efforts
demanded exploring other options with and without high-lysine mutants.
Several approaches were tried and critically examined for their merits and

S.K. Vasal (*)

CIMMYT, Texcoco, Mexico
e-mail: svasal12@yahoo.com

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 21
DOI 10.1007/978-81-322-1623-0_2, # Springer India 2014
22 S.K. Vasal

demerits. Different approaches were selected and emphasized in different

institutions. These will be briefly discussed, but emphasis will be given as
to why these approaches failed to produce desired results. CIMMYTs
approach has proven to be viable and has been successfully tried in
programs still maintaining interest in this particular maize. Selection of
appropriate strategy(ies) and breeding method(s) was the key to
CIMMYTs success. One-shot strategy as opposed to several individual
problem-related strategies was a must. Based on experience and available
information, the choice was to use a combination of two genetic systems
involving the opaque-2 gene and genetic modifiers of the opaque-2 locus.
At the very outset, there was a hope that this system can resolve complex-
ity of problems plaguing such materials. A series of steps were practiced
to implement this strategy and to start using it on a larger scale. Variation
in kernel modification had to be encountered and donor stocks had to be
built up initially. Some chance events will be mentioned which greatly
facilitated rapid development of QPM donor stocks. The next challenge
was to develop QPM germplasm adapted to different ecologies. A wide
array of breeding approaches was used to accomplish this needed task.
These will be fully discussed as to their relevance then and now.
Huge QPM germplasm volume was built through these varied efforts.
By mid- to late 1970s, improvement of QPM populations through inter-
national testing had already begun. Germplasm merging process was
initiated at this stage to form manageable number of QPM populations
and pools so that systematic improvement process could be started. At this
point, an important decision was also made to work in homozygous
opaque-2 backgrounds. The next turning point in QPM breeding efforts
was in mid-1980s when hybrid development was started. Combining
ability information and heterotic patterns of important classes of QPM
germplasm were established. Other aspects of hybrid development such as
inbred development, hybrid formation, and testing were initiated on a
modest scale. Even with a few hybrids formed and tested, the results were
quite encouraging. Several QPM hybrids had performance levels quite
similar or superior to better performing normal hybrids. The past 15 years
or so have witnessed much greater emphasis on QPM hybrid formation
and testing at CIMMYT. The international testing of QPM hybrids has
substantially increased, and many national programs are finding these
hybrids quite satisfactory in performance to the available commercial
hybrids. Several countries have released hybrids while others are on the
verge of releasing soon. Future strategies as well as traditional and modern
methodologies and tools that need to put in place will be discussed to have
broader QPM germplasm base available worldwide for breeding efforts.
One point, at the end, which I want to highlight is that support from
biochemical laboratory was an important factor for the success of QPM
work at CIMMYT. I consider QPM research as an excellent example of
interdisciplinary team effort at CIMMYT where biochemists and breeders
have worked in complete coordination to produce desired and acceptable
QPM products.
2 Breeding Challenges and Perspectives in Developing and Promoting Quality. . .
2.1 Introduction
Improving nutritional quality of agricultural crops is
a noble goal. Nutritional goals in general are difficult
and the success stories are relatively few. To achieve
good results requires long-term sustained
investments, patience, deep commitment, and perse-
verance in pursuing the desired goals. Improving
nutritional quality in cereal crops is particularly
important as the benefits can easily spread to
hundreds of millions of people in a most rapid
and effective manner without changing the tradi-
tional food habits. About maize, biochemists had
demonstrated several decades ago that maize protein
is nutritionally deficient because of the limiting
quantities of two essential amino acids: lysine and
tryptophan (Osborne and Mendel 1914). In the
absence of specific genes that could elevate protein
profile with respect to these amino acids, serious
research efforts by and large were lacking and lim-
ited to screening elite and bank maize accessions in
the hope of finding useful genetic variation for these
traits (Aguirre et al. 1969; Paez et al. 1969b;
Tello et al. 1965; Zuber 1975; Zuber and Helm
1975; CIMMYT 1970). As expected, variation was
encountered, but breeding efforts were not pursued
to make use of these materials in developing high
protein quality strains. Research initiatives had to
wait almost 50 years until three distinguished Purdue
researchers discovered high-lysine mutants such as
opaque-2 (Mertz et al. 1964) and floury-2 (Nelson
et al. 1965). The discovery of the biochemical
effects of these two mutants paved the way for
genetic manipulation in breeding to improve the
nutritional quality of maize endosperm protein.
Introduction of mutant genes into normal maize
brings a twofold increase in the levels of lysine and
tryptophan. Lysine-deficient zein or prolamine frac-
tion is reduced dramatically by about 50 %, while
other fractionsalbumins, globulins, and
glutelinsrich in lysine show a marked increase.
Most maize scientists envisioned this report with
great optimism and high hopes. Worldwide conver-
sion programs were initiated producing a wide
array of opaque-2 varieties and hybrids. These
high-lysine conversions were tested extensively in
the developed and the developing world. Yield and
agronomic performance of these materials did not
23
match the normal counterparts (Alexander 1966;
Harpstead 1969; Lambert et al. 1969; Gevers 1972;
Glover and Mertz 1987; Sreeramulu and Bauman
1970; Vasal 1975; Vasal et al. 1979, 1980, 1984a, b;
Bjarnason and Vasal 1992). In general, these
materials had 1015 % less grain yield, the kernel
appearance dull and chalky, slower drying, greater
kernel rots, and greater vulnerability to stored-grain
pests. The reasoning and causes behind these
problems are discussed in earlier publications
(Bjarnason and Vasal 1992; Vasal 1993, 2000,
2002; Prasanna et al. 2001). Thus in the very first
decade, this maize did not pass the test of its strength.
Frustration and declining interest was a consequence
of its lacking competitive performance. Funding
support was either reduced or completely with-
drawn. Most national maize programs in the devel-
oping countries reacted in a similar fashion.
Following mid-1970s, only a few institutes and
programs showed continuing interest and enthusi-
asm. CIMMYT had been one of them. The other
institutions and programs which sustained and
persisted in their breeding efforts were Purdue Uni-
versity, Crows Hybrid Seed Company in Milford,
Illinois, and the South African program at the Uni-
versity of Natal.
CIMMYTs work on quality protein maize
(QPM) is considered significant. It has achieved
scientific breakthrough in developing germplasm
products that are competitive and meet acceptance
of farmers and consumers preference (Vasal
2000). Several elements have contributed to this
success including sustained funding, strong admin-
istrative support at DG and directors level, readi-
ness to deploy alternate sound options in breeding
when the rest of the world was witnessing declining
interest, continuous evolving breeding methodo-
logies and strategies, and making constant changes
and adjustments as and when needed during differ-
ent phases of germplasm development.
2.2 Turning Points in Developing
Nutritionally Enhanced Maize
In developing nutritionally enhanced maize with
superior protein quality, the breeding program at
CIMMYT has evolved continuously. Breeding tac-
tics and strategies were modified or changed to
24
better options depending on the situation. As these
changes were made, the workload in protein labora-
tory and the ability to perform rapid and timely
analysis were given due attention (Villegas et al.
1992). Sequence of events in developing QPM
germplasm are discussed below in a brief manner.
2.3 Development and
Improvement of Soft Opaques
In the early period from 1966 to 1973, the emphasis
was on developing high-lysine soft endosperm
versions of normal materials. Initially, both opaue-
2 (o2o2) and floury-2 (fl2fl2) mutants were used
extensively, but very soon the use of floury-2 was
discontinued in the breeding program. Being semi-
dominant, the kernel expression and quality
depended on dosage of fl2 allele in the triploid
endosperm. Floury-2 conversion programs were
thus difficult to handle, and deployment of any
double mutant combination strategy was even
more difficult. In contrast, the opaque-2 mutant
had some obvious advantages. It was inherited in
a simple recessive manner, the conversion process
was easy, and no dosage effects were encountered
in the endosperm. Soft chalky phenotype associated
with the opaque-2 kernels was additional help as it
served as a marker and greatly facilitated the con-
version process. All potentially important maize
materials in the maize program were converted to
opaque-2 with varying numbers of backcrosses. A
few opaque-2 materials were also introduced from
other countries such as Thai opaque-2 composite,
Venzuela-1 opaque-2, and Ver.-Antigua opaque-2.
In addition, at least three broad-based opaque-
2 composites K, J, and I were formed to serve
lowland, mid-altitude, and highland agroecologies.
Population improvement efforts were undertaken in
some populations and population crosses were also
developed. It is fair to say that a larger share of the
resources was invested in developing and improv-
ing soft opaques. The materials so developed were
tested nationally and internationally in OMPT-11
trials. In the absence of hybrid program, no efforts
were made to develop and convert maize lines. This
was, however, attempted in many countries with
emphasis on hybrid maize as was the case with
S.K. Vasal
Brazil and Colombia. It may be interesting to
point out that partially modified kernels were
observed during the conversion process, but their
significance and importance were not realized and
appreciated until the first published report (Paez
et al. 1969a). In addition to large-scale research
effort on soft opaques, the program during this
period was also engaged in a small and insignificant
way to address some issues of exploratory nature.
These included screening bank accessions for
lysine and tryptophan content as well as developing
genetic isolation mechanisms to prevent contami-
nation in opaques. Double mutant combinations
of o-2 and fl-2 with waxy were also developed.
A beginning in modified opaques had already
begun considering the wisdom and vision of
Dr. Lonnquist and Dr. Asnani. By early to mid-
1970s, the problems plaguing soft opaques were
well recognized and documented in the developing
and the developed world.
2.4 Correcting First-Generation
Problems and Exploring
New Alternatives
It was mentioned earlier that soft opaques were
confronted with complex and interrelated
problems. It thus became imperative to consider
other alternatives in developing high-quality pro-
tein maize. Several new ideas emerged
suggesting options with and without the use of
mutant genes. In approaches not involving
mutant(s), three options were considered and
tried. These were recurrent selection for high
lysine in normal maize, selection for multiple
aleurone layers, and increasing the size of germ.
Recurrent selection merits consideration but only
limited use has been made of this approach. It
requires well-established laboratory facilities to
perform precise analysis. Other constraints in this
approach include lack of wide range in lysine
values, difficulty of transferring the improved
trait to other genetic backgrounds, and perhaps a
long time span in realizing lysine values close to
opaque-2 and with no certainty of enhanced
biological value. The size of germ can be
exploited favorably to enhance both protein
2 Breeding Challenges and Perspectives in Developing and Promoting Quality. . .
quantity and quality. It will, however, be difficult
to achieve lysine levels matching o-2 allele. The
selection process may affect other traits adversely
and may have a negative effect on shelf life of
corn. Double mutant combinations of o-2 and fl-
2 with waxy were also developed but had the
problem of unacceptable kernel phenotype.
In the second category, options involving
mutant genes, also three options were
investigated. These were search for new and bet-
ter mutants, developing double mutant
combinations, and using a combined o-2 and
genetic modifier approach. The search for new
mutants has continued, and a few additional
mutantsopaque-7 (Mcwhirter 1971), opaque-
6 (Ma and Nelson 1975), and floury-3 (fl3fl3)
(Ma and Nelson 1975)have been discovered.
Unfortunately, none of these mutants offer any
advantage over opaque-2 gene. The double
mutant combination su2o2 appeared promising
as it had vitreous kernels and protein quality
marginally better than opaque-2 gene alone
(Paez 1973). The kernels are, however, smaller
in size reflecting reduced grain yield of about
1525 % depending on the genetic background.
The yield gap can be narrowed down through
recurrent selection, but it is highly improbable
that the gap can be eliminated completely
(Vasal et al. 1980; 1984b; Vasal 1993, 2000,
2002). The double mutant o2fl2 has resulted in
vitreous kernels, but unfortunately it happens in
only rare backgrounds. In addition, the double
mutant shows effects on lysine, protein, and
light transmission (Paez et al. 1970). The com-
bined o2 and genetic modifier approach appeared
promising in altering kernel phenotype and to
remedy some of the defects associated with the
introduction of o2 gene. This approach has been
used widely in developing agronomically accept-
able germplasm.
Enough information and confidence in this
approach were already available when CIMMYT
maize researchers wanted to make a switch over
in the breeding program. Another important
point which I wish to make is that effort devoted
to developing soft opaques was not a complete
waste. It is from these materials we had to
look desperately for variation in modified kernel
behavior and got initial hints as to the types
25
of genetic backgrounds which will enhance
our chances of success in developing QPM
(Vasal 2000).
2.5 Genetic Modifiers and Their
Use as a Successful Strategy
Earlier several breeding options were mentioned
and briefly discussed. Unfortunately, most, if not
all, had shortcomings of different nature and
found no practical use in breeding program. The
approach that seemed to have promise involved
exploitation of genetic modifiers of opaque-
2 locus. It offered hope in improving phenotype
of soft opaques to hard and in remedying other
problems confronting these materials. CIMMYT
scientists viewed this approach with great opti-
mism and felt that it will have great application
in the future in developing high-quality protein
materials meeting producer, consumer, and
industrial acceptance. The genetic modifiers are
complex in their inheritance, the additive genetic
effects being more important than the dominance
effects. Also their expression varies with the
genetic background of the material in regard to
kernel modification, density, and biochemical
composition of the grain. Our own experience
also demonstrates that in some genotypes, pro-
tein quality may suffer a decline. Monitoring of
protein quality thus becomes an important com-
ponent in accumulating modifiers not affecting
quality. The new strategy thus brought successes,
but also new challenges unknown previously.
The progress was slow and frustrating in the
beginning as the starting point was zero or
so-called soft opaque-2 materials. Modifiers
also exert maternal influence thus complicating
the selection process. Negative influence on pro-
tein quality in some genotypes was disturbing
as protein quality had to be monitored
continuously and slowed down progress in
agronomic performance. Conversion program
remained no simpler. In fact, it became lengthy,
cumbersome, and difficult. The segregating
generations need to be handled much more care-
fully to avoid mistakes in selection. The breeding
work also had to depend on analyses provided by
the laboratory.
26
2.6 Development of QPM Stocks
Moving from soft opaques to hard endosperm
was highly critical if such materials were to be
accepted in developing countries. Any approach
under consideration should therefore involve
a change in kernel phenotype. Of the three
options at hand at that time, a combination
of opaque-2 gene and genetic modifiers was
considered most appropriate to rectify pheno-
typic appearance and other agronomic problems
affecting this maize. CIMMYTs choice of this
approach has proved to be a successful and a
viable strategy. No other option(s) used by
other institutes has demonstrated success of a
similar magnitude. Shift in strategy from soft to
hard endosperm was thus a major change but at
the same time an equally challenging task. There
was a lot of routine hard work, frustration in
encountering modified kernel variation, lack of
stability and reversion to soft texture in the ensu-
ing generation, and more disturbingly a decline
in protein quality in some instances. To deploy
this strategy on a massive scale required devel-
opment of white- and yellow-seeded QPM donor
stocks. Genetic variation for kernel modification
was encountered in several opaque-2 conversions
and opaque-2 populations under improvement.
Frequencies of modified kernels, however,
varied considerably among different genotypes.
Higher frequencies were observed in flint Cuban
and Caribbean germplasm as well as in Thai
opaque-2 composite and composite K. Modified
opaque-2 ears were selected independently from
several genetic backgrounds during the conver-
sion process and during maintenance and multi-
plication of opaque-2 materials. Selected kernels
were planted each generation separately from
each ear for several generations. This effort led
to the development of a large number of white
and yellow hard endosperm opaque-2 selfed
families. Later, white and yellow families were
recombined separately to give rise to white and
yellow hard endosperm opaque-2 composites,
respectively. This constituted the first approach
in developing QPM donor stocks. Genetic
mixing coupled with selection for modified ears
S.K. Vasal
and modified kernels of good protein quality was
practiced for 34 cycles.
The second approach involved intrapopula-
tion selection for modifiers in some populations
exhibiting a higher frequency of modified
kernels. Four tropical and one highland popula-
tion which met this criteria were chosen for this
purpose. These populations were composite K,
Ver.181-Ant.gpo.2  Venezuela-1 opaco-2, Thai
opaque-2 composite, PD (MS6) H.E.o2, and
composite I. The breeding procedure used was
controlled full-sib pollinations initially followed
by modified ear-to-row system suggested by
Lonnquist (1964). Between- and within-ear
selection for modification was practiced at all
stages.
Some unexpected chance events inspired con-
fidence and aided in a big way in deploying two
genetic systems strategy on a grand scale. This
was particularly true in a population cross of two
opaque-2 versions originating independently in
Thailand and Venezuela which exhibited a very
high frequency of mosaic modified kernels.
QPM donor stocks developed using above
mentioned approaches improved considerably
in the frequency of modifiers by mid-1970s.
At this point, it was decided to use these
materials as donor stocks as well as QPM
populations for further improvement using
appropriate schemes.
2.7 Expanded QPM Germplasm
Development Efforts
Choosing carefully a single approach/strategy
was critical in contrast to several independent
ones in tackling problems. Also any practical
strategy in use must address the problem of
yield penalty and phenotypic appearance of the
kernel. The strategy combined use of opaque-
2 gene and genetic modifiers was most appropri-
ate in this respect.
QPM germplasm development efforts were
not limited to a few genetic backgrounds. In addi-
tion, several different approaches and strategies
were put into action to develop a wide array of
germplasm products meeting the requirements
2 Breeding Challenges and Perspectives in Developing and Promoting Quality. . . 27

Table 2.1 Across-location performance of normal materials and corresponding tropical QPM germplasm, 1987

Grain yield (kg/ha)

Material Normal QPM QPM (% of normal)
Pool 23 5,405 5,330 98.6
Pool 24 5,706 5,457 95.6
Tropical high oil 5,733 5,170 90.2
Population 62 5,347 5,484 102.6
Population 65 5,255 5,369 102.2
Population 63 5,705 6,236 109.3
Source: CIMMYT

counterparts, some of the QPM versions have

Table 2.2 Mean grain yield of four highest yielding
QPM varieties, normal reference variety, and best local
checks at 11 locations, CIMMYT EVT 15B, 1988
Grain yield
(% of normal
Variety Grain yield (t/ha) ref. entry)
Across 8563 QPM 5.68 103.2
Capinopolis 8563 QPM 5.48 99.7
Iboperenda 8563 QPM 5.47 99.4
Tarapoto 8563 QPM 5.37 99.7
Suwan 8222 NRE 5.50 100.0
Best local check 5.36 97.5
Source: CIMMYT
for different agroecologies. The tactical strategy
was to go in a big way to develop huge germplasm
volume followed by merging and reorganization
of germplasm considering adaptation, maturity,
and kernel colors. This would help reducing
germplasm to a manageable level and permit
systematic handling. We are harnessing benefits
of this strategy, without which QPM may have
found only limited use in some countries.
In developing huge volume of QPM
germplasm, the thrust was on two principal
approaches. The first was to convert most of the
advanced unit populations representing tropical,
subtropical, and highland adaptation involving
different maturities, grain color, and texture.
A backcross-cum-recurrent selection procedure
was used for this purpose (Vasal et al. 1984b).
During the conversion process, emphasis was
placed on yield, kernel modification and appear-
ance, reduced ear rot incidence, rapid drying, and
other agronomic attributes. Compared to normal
performed quite similar in yield and agronomic
traits (Vasal et al. 1984a, b, 1993).
The second approach involved formation and
improvement of QPM gene pools. These pools
were developed using different procedures but
had homozygous opaque-2 background to facili-
tate selection and accumulation of genetic
modifiers. A modified half-sib system was used
for the improvement of these pools. A total of
seven tropical and six subtropical QPM gene
pools were developed and improved continu-
ously for varying numbers of cycles.
There was proof of continuing progress in
improving germplasm which provided clear-cut
evidence for others to see and appreciate
(Bjarnason and Vasal 1992; Bjarnason et al.
1998; Vasal 2000, 2002; Vasal et al. 1994).
Improvements were quite evident in grain yield
(Tables 2.1, 2.2, 2.3, 2.4, 2.5, and 2.6) and
in improving kernel modification (Tables 2.5,
2.6, 2.7, and 2.8) in several different genetic
backgrounds. The protein quality as well was
maintained during the selection process
(Tables 2.9 and 2.10).
2.8 Reorganization and New
Thrusts in QPM Germplasm
Management Strategy
Beginning mid-1970s and extending next 56
years, a huge volume of QPM germplasm was
developed that could meet germplasm needs of
several production environments in the tropics
and subtropics. At this point, it was realized
that continuing conversion program will serve
28 S.K. Vasal

Table 2.3 Superior white QPM hybrids tested across fifteen locations at El Salvador, Guatemala, and Mexico, 1998
Yield Tryptophan Ear Silking Plant
Pedigree (t/ha) Ear rot (%) (%) modification (days) height (cm)
CML142  CML146 6.48 3.7 0.096 2.0 55 242
CML159  CML144 6.39 4.3 0.100 1.6 56 230
(CLQ6203  CML150) 6.28 5.7 0.088 2.1 55 239
CML176
CML145  CML144 5.81 5.8 0.840 2.0 54 241
CML158  CML144 5.59 7.1 0.103 1.3 55 228
CML146  CML150 5.48 8.1 0.084 3.6 56 222
POZA RICA 8763 TLWD 5.34 12.0 0.095 2.8 54 230
Normal hybrid check 5.58 9.5 0.070 2.0 56 228
Local checks: HB-83, CB-HS-5G, H-59, XM7712, GUAYOPE

Table 2.4 Performance of superior tropical QPM hybrids across 41 locations in Latin America and Asia, 19992000
Grain yield Ear rot Silking Endosperma Tryptophan in
Pedigree (ton/ha) (%) (50 %) hardness sample
(CML-141  CML-144) 6.40 5.5 55 1.6 0.088
(CML-141  CML-144) CML- 6.39 6.2 55 1.7 0.081
142
(CML-142  CML-146) 6.28 6.3 55 2.2 0.100
(CML-142  CML-150) 6.20 7.8 55 2.0 0.089
(CML-142  CML-150) CML- 6.08 7.5 55 2.0 0.086
176
(CLQ-6203  CML-150) 5.80 7.2 55 2.3 0.090
(CML-144  CML-159) CML- 5.64 6.0 56 1.7 0.094
176
(CML-144  CML-159) (RE) 5.93 5.9 56 1.9 0.093
Local Check-1 5.95 7.6 55 1.9 0.050
a
Rating scale 15: 1 completely hard, 5 completely soft

Table 2.5 Comparison of original and latest cycle of selection in two QPM gene pools
Yield (kg/ Days to Plant height Soft cars Ear Endospermb
Material Cycle ha) flower (cm) (%) aspecta modification
Temperate  tropical QPM C0 4,437 59 212 66 4.0 4.2
(flint) C12 4,920 55 192 4 2.2 1.6
Temperate white QPM C0 3,445 55 204 34 3.5 3.7
C5 3,669 55 192 10 2.3 1.9
a
Rating scale 15: 1 excellent, 5 poor
b
Rating scale 15: 1 completely modified, 5 completely soft

no further useful purpose. Instead, perhaps it will
be more appropriate to accelerate progress by
switching over to working in homozygous
opaque-2 genetic backgrounds. Merging, consol-
idation, and reorganization of QPM germplasm
were done to reduce the number of QPM gene
pools and populations to a level so that these
could be handled in a more systematic way
and have some correspondence between pools
and populations. From the available QPM
2 Breeding Challenges and Perspectives in Developing and Promoting Quality. . . 29

Table 2.6 Agronomic characters of first and last cycles of selection in six QPM gene pool and populations
Endosperm
Pool/population Cycles of selection Grain yield (kg/ha) Days to silka Plant height (cm) modification
Population 62 QPM C0 4572 57.7 239 3.4
C5 4825 56.7** 225** 2.0**
Population 65 QPM C0 4277 57.5 235 3.1
C5 4406 55.3** 228 2.0**
Pool 23 QPM C1 4919 60.3 251 3.9
C19 5146 57.4** 227** 1.8**
Pool 24 QPM C1 5007 60.9 243 3.9
C19 5450** 58.6** 234* 2.4**
Pool 31 QPM C0 5834 65.9 211 3.0
C16 5880 65.0** 195** 1.6**
Pool 32 QPM C0 5883 66.3 210 3.1
C16 6172** 65.2** 200** 2.0**
Source: CIMMYT (Bjarnason, Short, Vasal, and Villegas)
*, **Significant at 5 % and 1 % probability levels respectively
a
Rating scale 15: 1 completely modified, 5 completely soft

Table 2.7 Improvement in kernel modification during different cycles of selection

Kernel modificationa
Material C0 C4 C9 L.S.D (.05)
Temperate  Trop. H.E.o2 (flint) 3.1 2.6 2.1 0.26
Temperate  Trop. H.E.o2 (dent) 3.0 2.5 2.2 2.20
Blanco Cristalino H.E.o2 3.7 2.8 2.3 0.20
Amarillo Dentado H.E.o2 3.2 2.6 2.4 0.43
a
Rating scale 15: 1 completely vitreous, 5 completely soft

Table 2.8 Mean kernel modification score, standard deviation, range, and Weinberg constant values of different
cycles of selection in yellow QPM grown in 1978
Cycle x kernel modification scorea Standard deviation Range Weinberg constant
C1 3.7 1.07 2.94.8 1.60
C4 2.9 0.92 2.33.7 1.57
C7 2.3 0.81 1.63.2 1.29
C9 2.2 0.91 1.23.5 1.23
a
Rating scale 15: 1 completely modified, 5 completely soft

germplasm, seven tropical and six subtropical

gene pools were formed and these are listed in
Table 2.11. In addition, six tropical and four sub-
tropical populations were developed (Table 2.12).
The handling of QPM gene pools and QPM
populations has been discussed in earlier
publications (Vasal et al. 1979, 1984a, b, 1997).
2.9 QPM Hybrid Development
and Testing
There have been several turning points in QPM
research at CIMMYT. This led to changing
emphasis in the program more than once in
30 S.K. Vasal

Table 2.9 Percent protein, tryptophan, and lysine in the whole grain of QPM pools
QPM gene pool Protein (%) Tryptophan in protein (%) Lysine in protein (%) Quality index
Pool 15 QPM 9.1 0.94 4.2 4.6
Pool 17 QPM 8.9 1.04 4.5 4.5
Pool 18 QPM 9.9 0.93 4.0 4.6
Pool 23 QPM 9.1 1.03 3.8 4.2
Pool 24 QPM 9.4 0.92 3.8 4.0
Pool 25 QPM 9.8 0.94 4.0 4.0
Pool 26 QPM 9.5 0.90 4.1 4.3
Pool 27 QPM 9.5 1.05 4.2 4.8
Pool 29 QPM 9.2 1.06 4.3 4.8
Pool 31 QPM 10.2 0.96 4.1 4.5
Pool 32 QPM 8.9 1.04 4.2 4.5
Pool 33 QPM 9.3 1.05 4.2
Pool 34 QPM 9.1 1.10 4.1 4.5

Table 2.10 Protein quality in different cycles of selection (endosperm analysis)

Material Cycle Protein (%) Tryptophan (%) Lysine (%)
CIMMYT H.E. o2 C0 8.0 0.78 2.90
C4 8.6 0.77 2.90
PD (MS) 6 H.E. o2 C0 8.5 0.78 2.50
C4 8.8 0.78 2.60

Table 2.11 QPM gene pools and their characteristics

QPM pool number Adaptation Maturity Seed color Seed texture
Pool 15 QPM Tropical Early White Flint-dent
Pool 17 QPM Tropical Early Yellow Flint
Pool 18 QPM Tropical Early Yellow Dent
Pool 23 QPM Tropical Late White Flint
Pool 24 QPM Tropical Late White Dent
Pool 25 QPM Tropical Late Yellow Flint
Pool 26 QPM Tropical Late Yellow Dent
Pool 27 QPM Subtropical Early White Flint-dent
Pool 29 QPM Subtropical Early Yellow Flint-dent
Pool 31 QPM Subtropical Intermediate White Flint
Pool 32 QPM Subtropical Intermediate White Dent
Pool 33 QPM Subtropical Intermediate Yellow Flint
Pool 34 QPM Subtropical Intermediate Yellow Dent

different years and at different stages of germ-
plasm development efforts. As discussed earlier,
some of these changes related to shifts in the use
of mutants or combined use of more than one
genetic system, changing emphasis from soft to
hard endosperm opaques, breeding approaches
and strategies, huge germplasm development
efforts initially and later consolidating into a
more definite number to satisfy germplasm
needs and permitting handling in a more
2 Breeding Challenges and Perspectives in Developing and Promoting Quality. . . 31

Table 2.12 QPM populations and their characteristics

Seed
Population number Name Adaptation Maturity Seed color texture
61 Early yellow flint QPM Tropical Early Yellow Flint
62 White flint QPM Tropical Late White Flint
63 Blanco Dentado-1 QPM Tropical Late White Dent
64 Blanco Dentado-2 QPM Tropical Late White Dent
65 Yellow flint QPM Tropical Late Yellow Flint
66 Yellow dent QPM Tropical Late Yellow Dent
67 Templado Blanco Cristalino QPM Subtropical Intermediate White Flint
68 Templado Blanco Dentado QPM Subtropical Intermediate White Dent
69 Templado Amarillo QPM Subtropical Intermediate Yellow Flint
70 Templado Amarillo Dent QPM Subtropical Intermediate Yellow Dent

systematic manner, and more importantly development including inbred development,

working in homozygous opaque-2 genetic
backgrounds. These constant changes and
adjustments were made to achieve rapid progress
in overcoming problems affecting QPM
materials and to emphasize competitive agro-
nomic performance of these materials to similar
or comparable normal counterparts.
Perhaps the next turning point was most impor-
tant and significant in QPM research efforts. This
was a new initiative related to introduction of
QPM hybrid research and development at
CIMMYT in 19841985 (Vasal 1986, 1987,
1994, 2002). This decision was particularly impor-
tant for QPM research as the use of inbred-hybrid
approach offered several advantages, including
seed purity, enhanced kernel modification and
added uniformity and stability, ease in monitoring
and maintaining protein quality, realizing
enhanced yield potential, addressing and circum-
venting problems more effectively and efficiently,
greater market acceptability, and more importantly
spurring seed industry growth. With the initiation
of hybrid development activities, combining abil-
ity information was generated, and heterotic
patterns of important classes of QPM germplasm
were established (Vasal et al. 1986; Vasal et al.
1993a, b). Inbreeding was introduced more
vigorously in population improvement activities
to improve inbreeding tolerance and to increase
the frequency of extracting superior inbreds
from such populations. Other aspects of hybrid
inbred evaluation, and hybrid formation were
initiated on a modest scale. Inbred and
hybrid performance also permitted formation of
synthetics based on combining ability information.
Even with a few hybrids formed and tested, the
results were very encouraging. Several QPM
hybrids had performance levels quite similar or
superior to better performing hybrids (Tables 2.3
and 2.4). The past 5 years or so have wit-
nessed much greater emphasis on QPM hybrid
formation and testing at CIMMYT. The interna-
tional testing of QPM hybrids has increased sub-
stantially and many national programs are finding
these hybrids quite satisfactory in performance to
the available commercial check hybrids. Several
countries have released hybrids based on
CIMMYT QPM lines while others are on the
verge of releasing soon. CIMMYT maize program
has also released 59 QPM lines in two separate
announcements in 1992 and 2003 (CIMMYT
1998, 2004).
Hybrid initiative turned out to be exceedingly
important for QPM efforts and success. Without
hybrid development efforts, it is likely that QPM
OPVs may not have made the same impact and
success stories. We are all aware that good sci-
ence is not free from difficulties, frustration,
and criticism. This should be viewed to generate
creativity, revisiting different approaches and
activities, and making constant adjustments for
efficient use of resources at all levels. Periodic
32
reviews help to de-emphasize some aspects of
the program while expanding others if necessary
and introducing new initiatives which were not
already in place. One good example will be
hybrid initiative in mid-1980s, which proved
extremely beneficial for QPM research and pro-
motional efforts. Without this initiative we per-
haps may not have realized full potential and
benefits of QPM. It is encouraging to state that
we are already harnessing the fruits of this impor-
tant decision. Many countries in recent years
have released directly CIMMYT-developed
hybrids and in a few cases are using CIMMYT
lines in combination with their own lines. The
outlook of QPM appears promising, and many
countries in the developing world are becoming
increasingly interested in QPM efforts.
2.10 Interdisciplinary Cooperation
and Some Important
Lessons Learned
During QPM development over the years, a
number of important lessons were learned
which have a bearing and relevance to similar
plant breeding situations in maize and other
crops especially where major gene(s) is used to
introduce some important attributes into the
elite germplasm. Invariably major genes with
drastic effects are associated with negative and
pleiotropic effects. To achieve success and
desired results will require accumulation of
favorable modifying alleles and will necessitate
the need to work in homozygous genetic
backgrounds contrary to commonly used back-
cross programs.
New initiatives are important and could make
a big difference and in some instances result in
breakthroughs. This is particularly true for those
breeding objectives which are important yet con-
troversial. Selection of breeding options and
strategies needs to be based on convincing evi-
dence and progress as was the case with the use
of combined opaque-2 and genetic modifiers
approach.
S.K. Vasal
Choosing carefully a single strategy/approach
is critical in contrast to several independent ones
in tackling multiple problems. The approach
used at CIMMYT in QPM research is an excel-
lent example to prove this point. Deploying the
right strategy of o2 and genetic modifiers, all
problems plaguing these materials were
addressed and good progress made on all fronts.
In problematic situations as was the case
with high-lysine mutants, the germplasm efforts
should not be limited to a few genetic
backgrounds. Also several different approaches
and strategies need to be put into action to develop
a whole array of germplasm products meeting
requirements for different agroecologies. The
tactical strategy in QPM research was to develop
a huge volume of germplasm followed by merg-
ing and reorganization of total germplasm into a
definite numbers of pools and populations that
were required for different ecologies. This
helped reducing germplasm volume to a man-
ageable level and also permitted more system-
atic handling through appropriate breeding
schemes. We are harnessing benefits of this
strategy, without which QPM may have found
its use in only limited environments and perhaps
in only a few countries.
QPM development required continuous
monitoring of protein quality and thus needed a
strong support from biochemical laboratory
(Villegas et al. 1992). Simple analytical
techniques were developed and used to analyze
large number of samples in a rapid and efficient
manner to provide results to the breeders in a
timely fashion to make right decision at right
time. This resulted in saving of resources
and manpower use. Collaboration between
biochemists and breeders was excellent, and
each group was aware of the needs, urgencies,
and work priorities of the other. Interdisciplinary
cooperation in QPM work at CIMMYT is per-
haps exemplary, and without the strong support
of laboratory, this work may not have attained
success that we are witnessing today. The
roles of other disciplines also cannot be
underestimated especially pathology, entomol-
ogy, and physiology which rendered services
2 Breeding Challenges and Perspectives in Developing and Promoting Quality. . . 33

Table 2.13 QPM releases involving CIMMYT germplasm

Name Institutions/country CIMMYT germplasm involved
SHAKTIMAN 1 DMR, India (CML 142, CML 150)
SHAKTIMAN 2 DMR, India (CML 176, CML 186)
HQ 2000 NMRI, Vietnam (CML 161, CML 165)
YunYao 19 Yunnan, China (CML 140)
Yun You 167 Yunnan, China (CML 194)
Qian 2609 Guizhou, China (CML 171)
Lu Dan 206 Shandong, China (P70)
Lu Dan 207 Shandong, China (P70)
Lu Dan 807 Shandong, China (P70)
Hybrid 2075 Sichuan, China (CIMMYT QPM populations)
Zhongdan 9409 CAAS, China (Pool 33 QPM)

Table 2.14 QPM releases in some countries 2.11 Renewed Interest and
No. of releases Emphasis on QPM Research
Country QPM OPV QPM hybrid Total
Mexico 5 21 26 Quality protein maize is a scientific breakthrough
India 1 2 3 and a great success story. Maize scientists feel
Brazil 3 3 proud of QPM research at CIMMYT and else-
Nicaragua 1 1 2 where. CIMMYT maize staffs have gained con-
Guatemala 1 1 fidence and strength in QPM research efforts and
El Salvador 1 1 are aspiring to do more to spread the benefits of
Honduras 1 1 this maize to more developing countries. Recent
China 5 5
releases and field days have been impressive
Colombia 1 1
(Tables 2.13, 2.14, 2.15, and 2.16). Expectations
Mozambique 1 1
of all of us are to have more area under this maize
Mali 1 1
in the developing countries in the future.
Uganda 1 1
Benin 1 1
Sasakawa Global 2000 under the leadership of
Burkina Faso 1 1 Dr. Norman E. Borlaug and several other staff of
Guinea 1 1 this organization has played a vital role in pro-
South Africa 1 1 moting QPM in several countries of Africa. The
Peru 1 1 success has revitalized QPM research at
Venezuela 1 1 CIMMYT and in many national programs around
Vietnam 1 1 the world.
Total 16 37 53 The past few years have witnessed lots of
QPM excitement. Hybrid performance of
CIMMYT-developed QPM hybrids has been
and support in evaluating QPM germplasm under very encouraging (Tables 2.3 and 2.4). It is time
different stresses. to think of plans for the future if widespread
QPM has been an excellent training ground benefits of this research are to be realized.
for maize researchers, especially postdoctoral There is a need to further strengthen QPM
fellows and visiting scientists. Researchers research at CIMMYT and in some of its regional
could be exposed to a whole array of activities programs. It is time that some of the strong NARs
going on in the maize program in addition to should also revive QPM research. Currently,
QPM. Many current CIMMYT staff had the priv- maximum use of CIMMYT germplasm is made,
ilege of working in QPM subprogram in the but this germplasm needs to be diversified and
initial stages of their careers. broadened at national and international level.
34 S.K. Vasal

Table 2.15 Recent QPM releases in some countries

Name Type Pedigree Country
HQ INTA-993 Hybrid (CML144  CML159) CML176 Nicaragua
NB-Nutrinta Open pollinated Poza Rica 8763 Nicaragua
HB-PROTICTA Hybrid (CML144  CML159) CML176 Guatemala
HQ-61 Hybrid (CML144  CML159) CML176 El Salvador
HQ-31 Hybrid (CML144  CML159) CML176 Honduras
Zhongdan 9409 Hybrid Pool 33  Temp QPM China
Zhongdan 3850 Hybrid China
QUIAN2609 Hybrid Tai 19/02  CML171 China
Yun Yao 19 Hybrid (CML140) China
Yun You 167 Hybrid (CML194) China
Lu Dan 206 Hybrid (P70) China
Lu Dan 207 Hybrid (P70) China
Lu Dan 807 Hybrid (P70) China
Hybrid 2075 Hybrid (CIMMYT QPM Populations) China
ICA- Hybrid (CML144  CML159) CML176 Colombia
Susumaa Open pollinated Across 8363SR Mozambique
Obatampaa Open pollinated Across 8363SR Mali
Obangainaa Open pollinated Across 8663SR Uganda
Obatampaa Open pollinated Across 8363SR Benin
BR-473 Open pollinated Brazil
BR-451 Open pollinated Brazil
AssumPreto Open pollinated Brazil
Obatampaa Open pollinated Across 8363SR Burkina Faso
Obatampaa Open pollinated Across 8363SR Guinea
QS-7705a Hybrid South Africa
GH-132-28a Hybrid P62, P63 China
INIA- Hybrid CML161  CML165 Peru
FONAIAP Hybrid (CML144  CML159) CML176 Venezuela
HQ-2000 Hybrid CML161  CML165 Vietnam
SHAKTIMAN-1 Hybrid (CML142  CML150) CML176 India
SHAKTIMAN-1 Hybrid CML176  CML186 India
In Mexico, 21 hybrids and 5 open pollinated varieties including. . .
44IC Hybrid CML142  CML116 Mexico
H-551C Hybrid CML142  CML150 Mexico
H-553C Hybrid (CML142  CML150) CML176 Mexico
H-519C Hybrid (CML144  CML159) CML170 Mexico
H-368EC Hybrid CML186  CML149 Mexico
H-369EC Hybrid CML176  CML186 Mexico
VS-537 C Open pollinated POZA RICA 8763 Mexico
VS-538 C Open pollinated ACROSS 8762 Mexico
2 Breeding Challenges and Perspectives in Developing and Promoting Quality. . .
Table 2.16 Some prominent QPM varieties and hybrids released in different countries
S.
no. QPM varieties/hybrids Countries
1 (CML144  159)  CML176 Nicaragua, Guatemala, El Salvador, Honduras, Mexico, Venezuela,
Colombia
2 CML176  CML186 India, Mexico
3 CML161  CML165 Peru, Vietnam, Guangxi (China)
4 (CML142  CML150)  CML176 India, Mexico
5 Across 83635R Mozambique, Mali, Uganda, Benin, Burkina Faso, Guinea
6 Poza Rica 8763 Nicaragua, Mexico
More researchers need to be trained who can
deploy QPM skills. Resources should be
mobilized for more QPM work, and future
strategies should make increasing use of modern
technologies and tools. QPM dissemination
should accompany extensive on-farm research
and demonstration of superior QPM varieties
and hybrids on farmers fields.
References
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Bjarnason M, Vasal SK (1992) Breeding of Quality Pro-
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lines (CMLS), CML1-CML497, pp. 159
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performance of normal and modified protein (opaque-
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changes protein composition and increases lysine con-
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opaque-2 maize kernels having different phenotypes.
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development of QMP hybrids. In: Anais do XV
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(1980) Genetic modifiers and breeding strategies in
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(1984a) Combined use of two genetic systems in
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protein maize-what is and how was it developed.
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Ross Publishing, Stroudburg, pp 241252
 Maize Protein Quality and Its
Improvement: Development of Quality 3
Protein Maize in India

M.L. Lodha

Abstract
The quality of maize protein is poor due to the deficiencies of two
essential amino acids, lysine and tryptophan, and excess of leucine.
However, the discovery of association of high lysine and tryptophan
with opaque-2 maize endosperm in 1964 opened up new vistas in improv-
ing maize protein quality. Consequently, many countries started develop-
ing maize varieties incorporating opaque-2 gene, and in the Indian Maize
Programme, three opaque-2 composite varieties were developed and
released for commercial production in the year 1970. Though the
nutritional superiority of opaque-2 maize over normal maize was very
well established, the newly developed high-lysine varieties could not
become popular because of agronomic and acceptability problems
associated with their soft chalky endosperm. To circumvent this problem,
efforts were initiated in 1971 itself towards developing hard-endosperm
opaque-2 maize lines/strains through the continuous process of vigorous
selection for kernel vitreosity and monitoring tryptophan/lysine for
maintaining protein quality. The modified opaque-2 maize with hard endo-
sperm and having high lysine and tryptophan is known as quality protein
maize (QPM). At present, in India we have one QPM composite and more
than nine QPM hybrid varieties released, and many more are in pipelines.
However, what is required is the popularization of these varieties with
farmers as well as consumers highlighting their nutritional significance,
especially to the vulnerable group, i.e. infants and preschool children and
pregnant and lactating mothers.

3.1 Introduction

In India, maize is the third most important food

M.L. Lodha
Ex- Professor and Head, Division of Biochemistry, Indian
Agricultural Research Institute, New Delhi 110 012, India
e-mail: mll_bio@yahoo.co.in
crop after rice and wheat. Its production has
reached 19.77 million tons, out of which about
25 % is used as human food, 12 % as animal feed
and 49 % as poultry feed (Dass et al. 2008). Thus,

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 37
DOI 10.1007/978-81-322-1623-0_3, # Springer India 2014
38
maize plays an important role in food and
nutritional security. Besides being the principal
source of carbohydrates and energy, maize like
other cereals is also the largest single source of
protein in the diet of the people for whom it is a
staple food. However, the nutritional quality of
maize protein is poor because of imbalanced
amino acid composition due to deficiencies of
two main essential amino acids, lysine and trypto-
phan, and excess of leucine. Alcohol-soluble pro-
tein fraction zein (prolamine), which is extremely
deficient in lysine and tryptophan, is the most
abundant protein (4060 %) in maize endosperm.
In whole grain of maize varieties, the protein
content ranges from 8.5 % to 13.6 %, whereas
lysine from 2.5 % to 3.6 % and tryptophan from
0.37 % to 0.67% of protein. In mature maize
kernel, endosperm accounts for 8085 % and
contributes as much as 80 % of total kernel pro-
tein. Embryo (germ) accounts for 810 % of total
weight and contributes 1520 % of total protein.
The niacin/tryptophan deficiency disease, pellagra,
caused by high concentration of leucine or high
leucineisoleucine ratio, is commonly associated
with maize-eating people. The deficiency is
recognized by 3 Ds: dermatitis, diarrhoea and
dementia.
The discovery of association of high lysine
and tryptophan with opaque-2 maize endosperm
by Mertz et al. (1964) opened up new vistas in
improving the protein quality of maize. The
opaque-2 (o2) single recessive gene, which is
located on chromosome 7, encodes a transcrip-
tional activator that regulates the expression of
alpha-zein genes (Schmidt et al. 1990). In the
presence of opaque-2 gene, the synthesis of 22
kD alpha-zein, which is extremely deficient in
lysine and tryptophan, is reduced and the synthe-
sis of non-zein protein fractions, particularly
glutelin, is increased in the endosperm resulting
in overall increase in the concentrations of lysine
and tryptophan in endosperm protein. In addi-
tion, there exists a highly positive correlation
between lysine concentration and protein synthe-
sis factor EF-1alpha, which is a lysine-rich abun-
dant protein and can be used as a biochemical
marker in the assay of lysine-containing proteins
(Habben et al. 1995; Moro et al. 1996).
M.L. Lodha
3.2 Essential Amino Acids and
Protein Quality
Proteins are an important constituent of our diet
and are required for (i) foetal development in
pregnancy and milk output during lactation, (ii)
growth in infants and children and (iii) mainte-
nance (replacing the wear and tear in tissues) in
adults. Amino acids are the building blocks of the
proteins. Food and tissue proteins contain 20
amino acids, of which eight (nine for infants)
are designated as essential amino acids, since
they cannot be synthesized in the body. The rest
of the amino acids are called non-essential as
they can be formed in the body. The proteins
needed by our body are supplied through the
diet we consume. The dietary proteins are broken
down into amino acids, absorbed as such and
used by the body to synthesize the proteins
needed for various functions like tissue building,
replacement of proteins depleted and synthesis of
various functional molecules.
The quality of dietary proteins depends on the
pattern of essential amino acids it supplies. The
best quality protein is one which provides essen-
tial amino acid pattern very close to the pattern of
the tissue proteins. The minimum amount of
essential amino acids required by infants is also
taken as reference pattern for defining the quality
of proteins. Milk and egg proteins, which are of
superior quality, also serve as reference protein.
As per the recommendations of the Joint FAO/
WHO Expert Consultation (1991), the essential
amino acid composition (mg/g protein) of refer-
ence protein for 25-year-old children is lysine
58, threonine 34, tryptophan 11 and methio-
nine + cysteine 25. As such, the first limiting
amino acid in cereal proteins is lysine, while
the second limiting amino acid is threonine/tryp-
tophan. Pulse proteins are rich in lysine but defi-
cient in sulphur-amino acids, mainly methionine.
Cereal or pulse proteins individually are, there-
fore, incomplete proteins. However, when
cereals and pulses are consumed together, they
have a mutually supplementary effect; deficiency
of an amino acid in one can be made good by an
adequate level in another. A mixed cereal-based
3 Maize Protein Quality and Its Improvement: Development of Quality Protein Maize in India
diet can meet the protein requirements of adults
and older children provided they consume
enough of the diet to meet their energy needs.
However, it is the vulnerable group of infants and
preschool children who need protein of superior
quality, and cereal-based diet or a mixed diet of
plant proteins cannot meet their protein
requirements adequately. For this group, the
requirement of protein per unit body weight is
also higher, the safe level of reference high-
quality protein for preschool children being
1.1 g/kg and for adults 0.75 g/kg body weight.
The nutritive value of food grain proteins,
depends upon their essential amino acid make
up as well as digestibility. For chemical evalua-
tion of protein quality, first and/or second limit-
ing amino acid in respective food grains is
determined. For example, in maize protein,
lysine is the first limiting amino acid and trypto-
phan is the second. The overall quality, i.e. the
nutritive value of food grain proteins is deter-
mined in a nitrogen-balance study with labora-
tory animals like growing albino rats. From this
study, biological value (BV) of proteins, which is
a percentage of N-retained by the experimental
animals out of N-absorbed from the diet, is
obtained. This test also gives an estimate of true
digestibility (TD) of proteins which indicates the
per cent N-absorbed from the diet. Net protein
utilization (NPU) value, a multiplication product
of TD and BV, gives a measure of per cent N-
retained out of total N-consumed. Utilizable pro-
tein (UP) value, a multiplication product of NPU
and per cent protein in the test food grain, is a
measure of both protein quality as well as quan-
tity. Another simple measure of the nutritive
value of proteins could be protein efficiency
ratio (PER), which indicates the gain in weight
Table 3.1 Amino acid composition of three opaque-2 composites and normal maize hybrid endosperm protein
Amino acid (g/100 g endosperm protein) Ganga-5 (normal)
Lysine 1.88
Tryptophan 0.35
Leucine 14.76
Protein (%) 8.5
Source: Singh and Koshy (1974)
39
of young animals per unit weight of protein con-
sumed. For this type of study, usually weanling
albino rats are used.
3.3 Development of Opaque-2
Maize Varieties and
Improvement in Protein
Quality
Soon after the discovery that opaque-2 gene
improves the contents of lysine and tryptophan in
maize endosperm, maize breeders all around the
world started incorporating this gene into their
elite maize varieties. In India also the work started
in 1966, and three opaque-2 maize composite
varieties, namely, Shakti, Rattan and Protina,
were developed under the auspices of All India
Coordinated Maize Improvement Project and
released for commercial production during 1970.
In the endosperm of these varieties, lysine and
tryptophan contents increased by about 100 %
and 160 %, respectively, while leucine content
decreased by about 35 % as compared to normal
maize hybrid Ganga-5 (Table 3.1).
The opaque-2 gene apparently acts as zein
suppressant in the developing endosperm. In
normal maize endosperm, zein accounts for
more than 50 % and glutelin for about 35 % of
the total protein. In opaque-2 endosperm, how-
ever, glutelin becomes the major fraction and
zein content is reduced drastically. In addition,
there are relative increases in albumin and
globulin fractions (Table 3.2), which are rich in
lysine and tryptophan.
The nutritional quality of normal and opaque-
2 maize protein has been evaluated using albino
rats, swine, children as well as adults.
Opaque-2
Shakti Rattan Protina
4.07 3.76 3.68
0.92 0.93 0.90
9.19 9.72 9.26
9.20 9.0 6.1
40 M.L. Lodha

Table 3.2 Protein fractionation of normal, soft opaque and hard-endosperm opaque-2 versions of Tuxpeno-1 maize
Fraction Normal Soft opaque-2 Hard-endosperm opaque-2
I (Albumin + Globulin) 6.2 20.6 15.5
II (Zein) 39.2 8.1 10.4
III (Zein-like) 19.7 10.7 16.2
IV(Glutelin-like) 13.6 18.5 21.4
V (Glutelins) 22.4 42.5 36.6
Source: Vasal (1994)

Table 3.3 Nutritive value of proteins of normal and opaque-2 maize

Protein value
expressed as % Casein Vijay (composite) Ganga-5 (hybrid) Bassi (local) Shakti (opaque-2 composite)
TD 100 92.1 92.8 93.7 95.2
BV 84.1 56.4 59.8 56.0 76.0
NPU 84.2 52.2 55.6 52.6 72.3
UP 71.38 5.75 6.56 6.94 8.44
Protein (%) 84.81 11.00 11.81 13.19 11.69
Source: Lodha et al. (1976b)
TD true digestibility, BV biological value, NPU net protein utilization, UP utilizable protein

circumference. The experiments were carried out

3.4 Rat-Feeding Experiments
The nutritional superiority of opaque-2 maize
over normal maize has been well established in
feeding trials with albino rats as experimental
animals. In a nitrogen-balance study, the BV of
opaque-2 maize (Shakti) protein was found to be
90 % of milk protein casein, while that of normal
maize (Vijay, Ganga-5, Bassi) protein was less
than 70 % of casein (Lodha et al. 1976b). The
NPU and UP values were also higher for opaque-
2 maize (Table 3.3). The protein efficiency ratio
(PER) value for opaque-2 maize has been found
to be 62110 % superior to that of ordinary
maize, the PER for opaque-2 maize being
2.32.8 and that for normal maize 1.31.6. Fur-
ther, 1 g of dietary opaque-2 maize resulted in
0.226 g weight gain in albino rats as against only
0.131 g with normal maize.
under the auspices of All India Coordinated Maize
Improvement Project, IARI, for a period of 6
months (November 1975May 1976) with
1830-month-old children from low-income
families. The children were fed on isocalorie
(405 Kcal) and comparable protein (about 10 g)
diets as a supplementary midday meal, to meet out
one-third of the recommended daily allowances.
The children fed on opaque-2 maize diet gained in
weight 8.3 % and 25 % more as compared to those
fed on skim milk and normal maize-based diets,
respectively. As regards height, children fed on
opaque-2 maize diet gained 29.4 % more com-
pared to normal maize. Similarly, children of the
milk group showed maximum gains in chest and
arm circumference, followed by opaque-2 maize
group and least gains were observed in normal
maize group (Table 3.4).

3.6 Therapeutic Uses of Opaque-2

3.5 Child-Feeding Experiments Maize

Opaque-2 maize has been proved to be a good Studies carried out at Universidad del Valle
supplement to the home diet of weanling and Hospital in Cali, Colombia, and at the National
young children in terms of their weight and height Institute of Nutrition (NIN), Hyderabad, have
gain and gain in mid-arm and chest shown that children severely ill with kwashiorkor
3 Maize Protein Quality and Its Improvement: Development of Quality Protein Maize in India 41

Table 3.4 Average gain in weight, height, chest and mid-arm circumference of children after 6 months of supplementary
feeding
Gain in
Feeding group Weight (kg) Height (cm) Chest circumference (cm) Mid-arm circumference (cm)
Controla 0.72 2.86 1.94 0.23
Milk 1.20 4.22 3.12 0.74
Normal maize 1.04 2.99 1.90 0.16
Opaque-2 maize 1.30 3.87 2.97 0.47
Source: Singh et al. (1980)
a
For control group, the data were recorded after 5.5 months

Table 3.5 Nutritive value of proteins of developing kernels of normal and opaque-2 maize

Ganga-5 (normal) Shakti (opaque-2)

Protein value expressed as % 25 days 35 days Mature 25 days 35 days Mature
TD 76.3 81.9 92.8 80.0 84.4 95.2
BV 76.3 70.8 59.9 86.5 78.0 76.0
NPU 58.2 58.0 55.6 69.2 65.7 72.4
UP 7.8 7.3 6.6 8.8 7.7 8.4
Protein (%) 13.4 12.5 11.8 12.7 11.7 11.7
Source: Gupta et al. (1978)

can be easily cured from the protein deficiency

disease if fed regularly on opaque-2 maize diet.
In addition, another study carried out at NIN with
dogs suggested that if the population having maize
as a staple food regularly consumes opaque-
2 maize diet, then the incidence of pellagra could
be prevented.
3.7 Protein Quality of Immature
Kernels
In addition to their direct use after roasting or
boiling, immature kernels from green ears are
used in preparing many recipes. With the devel-
opment of the kernels, the TD of the protein is
improved; however, the BV decreases in both
normal and opaque-2 maize. The BV values
being equal (76.3 and 76.0), immature normal
maize kernels harvested at the early-dough
stage (25 days post-pollination) were found to
be as good as mature opaque-2 maize kernels
from their BV point of view (Table 3.5).
3.8 Problems Confronting Chalky
Opaque-2 Maize and
Development of Modified
Opaque-2 Maize
Though the nutritional superiority of opaque-
2 maize over normal maize has been very well
established, the opaque-2 maize varieties could
not become popular all over the world due to the
following agronomic and acceptability problems
associated with soft and chalky endosperm:
1015 % lower grain yield, poor kernel appear-
ance, high susceptibility to ear rot and stored
grain pests (Singh et al. 1974; Vasal 2000).
Fortunately, during the process of converting
normal maize populations to their opaque-
2 versions, partially hard endosperm (i.e. vitreous)
or modified grains had been observed by many
researchers including breeders at CIMMYT in
Mexico. Separation of such grains began as early
as in 1969 (Prasanna et al. 2001). At CIMMYT,
under the dynamic leadership of Dr. S.K. Vasal, and
at the University of Natal, various endosperm
42
modifier genes were identified that could
favourably alter the grain characteristics, thereby
overcoming an important obstacle in the populari-
zation of high-lysine opaque-2 maize (Sofi et al.
2009). The mechanism by which the endosperm
modifiers change the grain structure from chalky
to vitreous in modified opaque-2, is not clearly
understood. The modified opaque-2 maize with
hard endosperm is known as quality protein maize
(QPM). Thus, the term QPM now refers to maize
homozygous for the o2 allele, with increased lysine
and tryptophan contents but without the negative
secondary effects of a soft and chalky endosperm.
The QPM essentially has about twice the levels of
lysine and tryptophan than normal maize and also
increased levels of histidine, arginine, aspartic acid
and glycine, but reduced level of leucine.
In the Indian Maize Programme, though the ini-
tial emphasis was on the development of soft-
endosperm opaque-2 maize varieties, gradually the
emphasis shifted to developing modified opaque-
2 maize strains with hard endosperm. The work
was initiated during kharif (rainy season) of 1971,
and a large number of inbred lines of varying
vitreosity were selected from the three opaque-
2 composites, Shakti, Rattan and Protina. These
lines were subjected to vigorous selection for kernel
vitreosity by growing them in Delhi during kharif
(rainy season) and during rabi (winter season) in
Hyderabad for 3 years (19721974). After each
harvest, the selection was made based on kernel
vitreosity and tryptophan content. In each cycle of
selection, semi-opaque and semi-normal kernels
were sown separately. Based on light transparency,
the kernels were classified as opaque (O), 100 %
opaque with no light transmission; semi-opaque
(SO), partial light transmission; semi-normal (SN),
over two-third of the kernel from the crown is vitre-
ous; and normal (N), nearly vitreous kernels with
slight spotting or cloudiness. The development of
hard-endosperm inbred lines of opaque-2 maize
over five cycles of selection and their analyses
showed that protein content, 100-kernel weight and
kernel density increased with the increase in kernel
vitreosity; however, tryptophan (as % of endosperm
protein) decreased slightly. There was variability in
tryptophan content suggesting a possibility of
selecting hard-endosperm opaque-2 strains with
M.L. Lodha
high tryptophan (more than 0.7 % in endosperm
protein) and nearly normal-looking vitreous type of
kernels (Lodha et al. 1976a; Singh et al. 1974, 1985).
Utilizing the selected hard-endosperm opaque-
2 inbred lines, a SO/SN composite was developed
which was subjected to extensive biochemical and
nutritional studies. Later on, some of these lines
were used by the Directorate of Maize Research
(DMR) in developing a QPM composite, Shakti-1
(0.92 % tryptophan), which was released in 1997.
The major emphasis was, however, given on
the development of QPM hybrids by practically all
the breeders. CIMMYT-developed QPM lines
along with their own lines were mostly involved
in two-parent QPM hybrids. At present, in India
we have one QPM composite (Shakti-1) and
more than nine QPM hybrid varieties, including
Shaktiman and HQPM series and Vivek QPM 9.
The QPM version of extra-early Vivek Hybrid 9,
Vivek QPM 9, was developed by following
marker-assisted selection (MAS) and had been
released for commercial cultivation in 2008. Phe-
notypically, the kernels of the QPM hybrid are as
vitreous as those of its normal maize counterpart
(Fig. 3.1). This QPM hybrid contains 4.19 % lysine
and 0.83 % tryptophan in its endosperm protein
as against 3.25 % lysine and 0.59 % tryptophan
in its normal counterpart (Gupta et al. 2009).
The QPM hybrids, Shaktiman-1, Shaktiman-
2, Shaktiman-3 and Shaktiman-4, respectively,
contain 1.01 %, 1.04 %, 0.73 % and 0.93 %
tryptophan in their endosperm proteins. The pro-
tein in the endosperms of these QPM hybrids
ranges from 9.3 % to 9.9 %. Similarly, in the
QPM hybrids, HQPM-1, HQPM-5 and HQPM-7,
which respectively contain 0.94 %, 0.76 % and
0.72 % tryptophan, protein in the endosperms
ranges from 9.36 % to 9.80 %.
3.9 Chemical and Biological
Evaluation of Modified
Opaque-2 Maize Protein
Quality
As a result of selection for modifiers, several
changes occur in physical and biochemical
characteristics of the modified opaque-2 maize
3 Maize Protein Quality and Its Improvement: Development of Quality Protein Maize in India 43

Fig. 3.1 Normal and QPM

version of Vivek hybrid 9
(Courtesy: Dr. H.S. Gupta,
IARI, New Delhi)

Table 3.6 Nutritive value of proteins of different categories of modified opaque-2, chalky opaque-2 and normal maize

Vijay SO/SN composite kernel categories (modified opaque-2) Shakti

Protein value expressed as % (normal) 25 % opaque 50 % opaque 75 % opaque (opaque-2)
TD 92.7 97.1 95.5 94.5 94.4
BV 60.6 72.2 73.1 73.3 78.1
NPU 56.2 70.1 69.8 69.2 73.7
UP 5.41 8.28 8.20 8.00 7.97
Protein (%) 9.63 11.81 11.75 11.56 10.81
Source: Gupta et al. (1979b)

kernels. Based on light transparency, modified
opaque-2 maize kernels are classified into the
following categories: vitreous (nearly normal),
25 % opaque75 % vitreous, 50 % opaque50 %
vitreous, 75 % opaque25 % vitreous and
completely opaque. The vitreous fraction of
endosperm contains more protein but less lysine
and tryptophan than the opaque fraction (Lodha
et al. 1976a). With the increase in kernel
vitreosity, protein and leucine contents increase,
while lysine and tryptophan contents decrease
in the endosperm. This is because albumin,
globulin and glutelin fractions decrease, while
the zein fraction increases in the endosperm
with the increase in kernel vitreosity (Gupta
et al. 1979a). Although chemical analysis
showed differences in endosperm protein qual-
ity of various categories of modified opaque-2,
rat-feeding experiments showed more or less
similar nutritional quality of the grain protein
based on UP values, as described below.
Nitrogen-balance study carried out with
albino rats utilizing three kernel categories of
modified opaque-2 with hard endosperm (SO/
SN composite), viz. 25 % opaque, 50 % opaque
and 75 % opaque; normal composite Vijay and
Shakti opaque-2 showed about 20 % higher BV
for different categories of modified opaque-2 as
compared with normal maize (Table 3.6).
Although the BV of Shakti opaque-2 was slightly
better (6.68.2 %) than that of various categories
of modified opaque-2, the UP values of Shakti
and three categories of modified opaque-2 were
not significantly different from one another. The
change was mainly because of higher protein in
different kernel categories of modified opaque-
2 composite in comparison to Shakti composite,
which has got chalky endosperm (Table 3.6).
Chalky opaque-2 and modified opaque-2 maize
also supported better liver growth compared to
normal maize in albino rats. A similar trend was
also observed for protein per cent in liver.
44
Table 3.7 Protein distribution in the endosperm of normal, chalky opaque-2 and modified opaque-2 maize
Per cent of total recovered protein
Protein fraction Vijay (normal) Shakti (chalky opaque-2)
Albumin 3.56 13.92
Globulin 4.75 10.22
Prolamine (zein) 43.33 10.22
Glutelin 43.04 51.68
Residue 5.32 13.96
Source: Gupta et al. (1980)
So far as distribution of protein fractions in
the endosperm is concerned, albumin content
was more or less equal in chalky opaque-2 -
composite (Shakti) and modified opaque-2 with
hard endosperm (SO/SN composite), and it was
fourfold higher than in normal maize Vijay. The
content of globulin was also twofold higher in
chalky and 1.5-fold higher in modified opaque-
2 compared to normal maize. However, the con-
tent of zein was drastically reduced in opaque-
2 types, being 76 % lower in chalky opaque-
2 and 58 % lower in modified opaque-2. The
content of glutelin was about 20 % higher in
opaque types compared to normal maize
(Table 3.7).
3.10 Child-Feeding Experiments
Nutritional superiority of QPM over normal
maize was also demonstrated in a child-feeding
experiment. In a 6-month study carried out with
preschool children at Rajendra Agricultural Uni-
versity, Pusa, gains in weight and arm circumfer-
ence were, respectively, about 30 % and 100 %
higher for Shakti-1 (QPM composite)-fed group
compared to the group fed with normal maize
(DMR 2001).
3.11 Genetic Systems and Need for
Monitoring Protein Quality of
QPM
There are three distinct genetic systems which
are involved in the development of QPM:
(i) recessive homozygous allele of the opaque-
2 gene, (ii) modifiers for endosperm hardness and
M.L. Lodha
SO/SN composite (modified opaque-2)
14.58
6.95
18.06
52.79
7.62
(iii) amino acid modifiers which affect the
relative levels of lysine and tryptophan in the
endosperm (Gupta et al. 2009).
The recessive mutant allele of the opaque-
2 gene as described earlier is the first and most
important component. The second distinct
genetic system is comprised of the alleles of
endosperm hardness modifier genes which con-
vert the soft/opaque mutant endosperm to a hard/
vitreous endosperm with little loss of protein
quality. It has been shown that increased levels
of gamma-zein contribute to the recovery of a
hard-endosperm phenotype as the modified
opaque-2 (QPM) grains have approximately dou-
ble the amount of gamma-zein in the endosperm
relative to the o2-only mutants. These modifiers
along with the o2 mutant allele can be initially
selected by using a rapid and low-cost method of
selection, whereby light is projected through the
vitreous grains or blocked by the opaque grains,
respectively. Thus, when the kernels are screened
against light, varying degrees of modification,
ranging from completely vitreous to fully
opaque, may be observed. As shown in Fig. 3.2,
varying degrees of kernel modification may also
be observed in QPM ears.
Two genetic loci which affect the modifica-
tion of the endosperm hardness in o2o2
backgrounds have been mapped to the long arm
of chromosome 7, and interestingly one endo-
sperm modifier locus maps near a gamma-zein
gene gzr1. The third genetic system critical to a
QPM breeding programme is comprised of a
distinct set of amino acid modifier genes which
affect the relative levels of lysine and tryptophan
content in the grain endosperm. The lysine level
in normal and QPM maize average 2.0 % and
4.0 % of total protein in whole grain flour,
3 Maize Protein Quality and Its Improvement: Development of Quality Protein Maize in India
Fig. 3.2 Varying degree
of kernel modification in
QPM ears, ranging from
completely modified to
very low level of
modification (from left to
right) (Courtesy: Dr. B.M.
Prasanna, CIMMYT,
Kenya)
respectively, but range across genetic
backgrounds from 1.6 to 2.6 in normal maize
and 2.74.5 % in their o2-converted counterparts
(Moro et al. 1996). Since QPM looks like normal
maize (Fig. 3.1), there is a need to monitor lysine
or tryptophan levels in its endosperm continu-
ously during the breeding process.
After initial screening of QPM materials
against light, the endosperm samples of selected
materials having vitreous kernels are subjected to
chemical analysis for lysine/tryptophan and pro-
tein. Because of the existence of a relationship of
approximately 14 in tryptophan and lysine in
maize endosperm protein and also because of
simplicity of its estimation, initially the trypto-
phan content may be used as a single parameter
for protein quality evaluation. Because of its sim-
plicity and rapidity, a colorimetric method in
which a single-step papain hydrolysis is utilized
for protein solubilization is used more extensively
for screening maize germplasm for tryptophan
content (Villegas and Mertz 1971). With this
method, it is possible to analyse up to 75 samples
each day. Subsequently, the lysine determination
is performed only on those samples selected as
having high tryptophan values or when lysine
value in addition to tryptophan is desired. Because
of its simplicity and rapidity, a colorimetric
method modified by Villegas (CIMMYT), which
45
also utilizes a single-step papain hydrolysis for
protein solubilization, is followed extensively for
screening maize germplasm for lysine content
(Villegas and Mertz 1971). By this method up to
eight samples can be processed simultaneously,
and it gives an estimate of available lysine. In
addition, since the concentration of lysine in the
maize endosperm is highly correlated with the
content of a single non-zein, protein synthesis
factor EF-1 alpha, assay of its content by ELISA
also provides a reliable index of lysine in endo-
sperm (Habben et al. 1995; Prasanna et al. 2001).
Once a QPM variety reaches a final stage of its
release for commercial production, it is advisable
to analyse it for complete amino acid profile and
subject it to animal-feeding experiments for
biological evaluation of its nutritive value.
3.12 Improving Nutritive Value
of Normal Maize and QPM
by Amino Acids and Pulse
Supplementation
The protein quality of ordinary maize and even
of hard-endosperm modified opaque-2 maize can
be improved by supplementation with limiting
essential amino acids or pulses because of the
complementary nature of essential acids present in
46
pulses and cereals. The UP value, which takes into
account both protein quality as well as quantity,
showed substantial improvement with amino acid
supplementation as well as by addition of pulse to
normal and modified opaque-2 diet. Studies car-
ried out by Gupta et al. (1979b) with albino rats
showed that supplementation with lysine alone in
case of modified opaque-2 and with lysine + tryp-
tophan in ordinary maize improved substantially
the nutritive value. When modified opaque-2 was
supplemented with 0.22 % L-lysine HCl, its UP
value increased from 6.6 % to 8.3 %, showing
thereby about 25 % improvement in nutritive
value of protein. In the case of Vijay normal
maize, the UP value increased from 6.1 % to
8.2 % (about 34 % improvement) when it was
supplemented with 0.45 % L-lysine HCl + 0.06 %
DL-tryptophan. At the same time, synergistic
effect to the order of 25.0 % and 22.0 % has
been observed in the case of normal maize and
modified opaque-2 maize, respectively, when
these were supplemented with chickpea pulse
(7:3 on protein basis).
3.13 Future Strategies
The nutritional superiority of Quality Protein
Maize (QPM) over normal maize is well
established. QPM can become a boon to young
children, especially to those suffering from pro-
tein malnutrition. More so, because its consump-
tion does not require any change in traditional
food habits. However, there is a need to popular-
ize QPM varieties with farmers as well as
consumers, highlighting its nutritional impor-
tance so that its cultivation gains the required
momentum. To achieve this objective, it is also
important that farmers get remunerative price for
their produce. There is also a need to highlight
that QPM is a product of conventional breeding,
and no genetic engineering is involved in its
development. The produce may be utilized in
the midday meal programme of school children.
In addition, value addition is also required and
efforts are needed to popularize already devel-
oped QPM products, for example, infant food,
M.L. Lodha
health food, snacks and savoury items, conve-
nience food and specialty food (Singh 2001).
This is going to benefit specially the vulnerable
groups, i.e. infants, preschool children, pregnant
and lactating mothers and elderly people. From
the poultry industry point of view, concerted
efforts are required towards developing maize
rich in amino acid methionine.
The lysine content of presently available
QPM varieties is below 5 mg/100 mg protein,
the FAO/WHO recommended minimum level
for human diets. Therefore, by exploiting
variability for lysine content in QPM germplasm,
it may be possible to select economically viable
genotypes with lysine content exceeding the
recommended level. Moreover, it is also impor-
tant to establish as to how the modification of
opaque-2 locus by modifier genes is affected at
biochemical and molecular level. This should
help understand the mechanism of action of mod-
ifier genes, the expression of which is affected by
genetic and environmental factors.
3.14 Conclusion
An important goal of plant breeding programmes
has been improvement in nutritive value, in par-
ticular amino acid profile, of food grains. The
biggest success was achieved when high-lysine
opaque-2 mutant of maize was identified, and
subsequently QPM developed by exploiting
genetic modifiers of opaque-2 locus. QPMs are
the mutants that have a hard, vitreous endo-
sperm; a normal yield; and a high nutritional
quality. Since, there exist a lot of variability for
lysine/tryptophan content in the germplasm, and
breeding and development of QPM varieties by
accumulating the modifiers being a multistep
process, monitoring of tryptophan/lysine level
at each step is required as QPM kernels cannot
be easily distinguished visually from normal
maize kernels, and chemical analysis is the only
way. Thus, establishment of a few well-equipped
biochemical laboratories to provide rapid and
reliable analyses would be able to give a desired
boost to the ongoing QPM programme.
3 Maize Protein Quality and Its Improvement: Development of Quality Protein Maize in India
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Dass S, Jat ML, Singh KP, Rai HK (2008) Agro-economic
analysis of maize-based cropping systems in India. Ind
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DMR (2001) Production technology of quality protein
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Research, Pusa Campus, New Delhi
Gupta HO, Lodha ML, Singh J, Mehta SL (1978) Protein
quality of normal and opaque-2 maize at different
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(1979a) Nutritional evaluation of hard endosperm
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27:390392
Gupta HO, Lodha ML, Mehta SL, Rastogi DK, Singh J
(1979b) Effect if amino acid(s) and pulse supplemen-
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27:787790
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(1980) Changes in minerals, proteins and amino acids
in hard endosperm opaque-2 Zea mays during devel-
opment. Ind J Expt Biol 18:14191422
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BA (1995) Elongation factor-1 alpha concentration is
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SL, Singh J (1976b) Nutritive value of normal and
opaque-2 maize. Curr Sci 45:287289
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Quality protein maize. Curr Sci 81:3081319
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regulatory gene opaque-2 encodes a protein with a
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 Molecular Interventions for Enhancing
the Protein Quality of Maize 4
Yogesh Vikal and J.S. Chawla

Abstract
Maize, one of the three most popular cereal crops of the world, globally
contributes 15 % of the protein and 20 % of the calories derived from food
crops in the worlds diet. However, cereals do not provide a nutritionally
balanced source of protein. For nutritional security, it is necessary to adopt a
genetic enhancement strategy in which essential amino acids are either
incorporated or increased in grain protein to alleviate hunger, increase
income, and improve livelihood. Quality protein maize (QPM) is having
high nutritive value of endosperm protein with opaque2 (o2) mutation
leading to 60100 % increased content of lysine and tryptophan. The lysine
value of o2 maize is 2.54.0 g/100 g of endosperm protein, which is more
than twice that of the normal maize (1.3 g lysine/100 g protein). International
Maize and Wheat Improvement Center (CIMMYT), Mexico, played a
significant role in the development of QPM maize. The breeding of QPM
involves three genetic systems: (i) the recessive mutant allele of the o2 gene,
(ii) the endosperm hardness modifier genes, and (iii) the amino acid modifier
genes influencing free amino acid content in the endosperm. Due to recessive
nature of the o2 gene, complex action of modifier genes, and presence of
amino acid enhancer genes, the use of DNA marker-assisted selection (MAS)
accelerated the selection efficiency and expedited the development of new
QPM cultivars. Using a combination of MAS and phenotypic selection
techniques, a single cross, short duration Vivek QPM 9 hybrid was developed
and released in 2008 by Vivekananda Parvatiya Krishi Anusandhan
Sansthan, Almora, India. Alternatively, manipulating the plant lysine meta-
bolic pathway provides possible enzyme targets for genetic engineering to
increase free lysine content in corn grain. Furthermore, RNA interference

Y. Vikal (*)
School of Agricultural Biotechnology, Punjab
Agricultural University, Ludhiana, Punjab, India
e-mail: yvikal@hotmail.com
J.S. Chawla
Department of Plant Breeding and Genetics, Punjab
Agricultural University, Ludhiana, Punjab, India

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 49
DOI 10.1007/978-81-322-1623-0_4, # Springer India 2014
50 Y. Vikal and J.S. Chawla

(RNAi) has been used to specifically suppress -zein production in transgenic

corn, resulting in a doubling of the lysine content of corn grain. QPM is likely
to gain wider acceptance if QTLs for kernel modification, and enhancers for
amino acids are fine mapped to develop markers to follow MAS for vitreous
kernels and high levels of lysine. However, the major constraints in adoption
of QPM hybrids are contamination with normal maize pollen in field,
resulting in erosion of the trait in farmer-saved seed system. It is essential
to give training on good seed production practices to the local communities
and development of linkage between the seed producers, farmers, and the
industry for sustainable higher nutritional benefits of QPM in the long term.

4.1 Introduction For nutritional security, it is necessary to adopt a

Maize (Zea mays L.) is one of the worlds three
most popular cereal crops. It is a major cereal crop
for both livestock feed and human nutrition
worldwide and of great economic importance not
only as a grain and fodder crop but also as an
industrial raw material. Maize ranks next to rice
and wheat with respect to area and grain produc-
tion. India is the fifth largest producer of maize in
the world contributing about 3 % of the global
production. In India, maize was grown on an area
of 8.33 million hectare with productivity of 2.4
tonnes/hectare in the year 20102011. The
demand for cereal grains will continue to increase
as a consequence of the expanding human popu-
lation, which could add >1.5 billion people by the
year 2025 (Lutz 2001). However, cereals do not
provide a nutritionally balanced source of protein.
Protein-containing foods are necessary for the
rapid growth of children (Millward and Rivers
1989), and in some countries, maize is a primary
weaning food for babies.
From nutritional perspective, the protein of nor-
mal maize is deficient in the essential amino acids,
lysine and tryptophan (Bhatia and Rabson 1987).
Protein deficiency, especially in children, causes
kwashiorkor, a potentially fatal syndrome
characterized by initial growth failure, irritability,
skin lesions, edema, and fatty liver. Nutritional
deficiency is of concern, particularly for people
with high protein requirements, e.g., young chil-
dren, pregnant and lactating women, and patients.
genetic enhancement strategy in which essential
amino acids are either incorporated or increased
in grain protein to alleviate hunger, increase
incomes, and improve livelihoods.
4.2 Maize Protein
Maize kernel consists of endosperm, embryo, and
the outer seed coat. Both the embryo and endo-
sperm contain proteins but differ substantially in
the content and the quality of the protein. The
endosperm accounts for 8085 % and the embryo
810 % of the total kernel dry weight. The endo-
sperm of maize has two district regions. The aleu-
rone layer, the outermost layers, is rich in
hydrolytic enzymes. The starch-rich endosperm
is present within the aleurone layer, which is
vitreous (Fig. 4.1). The protein fraction constitutes
only 810 % of the endosperm, while starch
accounts for about 70 % of the kernel (Lawton
and Wilson 1987; Prasanna et al. 2001). The vari-
ous fractions of endosperm protein are albumin
3 %, globulin 3 %, zein (prolamine) 60 %, and
glutelin 34 %, whereas the embryo protein largely
consists of albumins (60 %).
The zein proteins found in vitreous region form
insoluble accretions called protein bodies in the
lumen of rough endoplasmic reticulum and
toward maturation are densely packed between
starch grains (Gibbon and Larkins 2005). Based
on their solubility, genetic properties, and the
4 Molecular Interventions for Enhancing the Protein Quality of Maize
Fig. 4.1 Structure of maize kernel (Source:http://www.
public.iastate.edu/~becraft/Endosperm.htm)
apparent molecular masses, zeins have been clas-
sified into -zein (22 and 19 kDa), the most abun-
dant; -zein (14 kDa); -zein (27 and 16 kDa); and
-zein (10 kDa) (Wilson et al. 1981). The zein
fraction is rich in cysteine, while and
fractions are rich in methionine. The zein fraction
in normal maize generally contains a higher pro-
portion of leucine (18.7 %), phenylalanine
(5.2 %), isoleucine (3.8 %), valine (3.6 %), and
tyrosine (3.5 %) but smaller amounts of other
essential amino acids such as threonine (3 %),
histidine and cysteine (1 %), methionine (0.9 %),
and lysine (0.1 %) and is devoid of tryptophan.
The non-zein protein fraction is balanced and rich
in lysine and tryptophan. The high proportion of
zeins in the endosperm is the primary reason for
the poor protein quality of maize (Vasal 2000).
Protein supplementation to correct such
deficiencies is costly and wasteful of energy in
animal nutrition and is not feasible for most devel-
oping countries, which rely on cereal for human
consumption. Therefore, increasing the amounts
of protein-bound essential amino acids depends
on improving cereal storage proteins.
51
4.3 High-Lysine Mutants in Maize
Several mutations, both spontaneous and
induced, have been recognized that affect the
amino acid composition of maize endosperm.
In the early 1920s, opaque2 (o2) mutation was
first described by Jones and Singleton; but later
on in 1963, researchers at Purdue University,
USA, discovered that opaque2 (o2) increases
grain proteins nearly twice as those of normal
maize proteins (Mertz et al. 1964). In fact,
maize homozygous for the recessive o2 allele
has substantially higher lysine (>69 %) and
tryptophan content compared to normal maize.
Nelson et al. (1965) discovered another muta-
tion, the floury2 gene, which in homozygous
form also increases lysine and tryptophan
levels in maize. Later on, a large number of
mutants were discovered, which fall into three
categories: the recessive mutants opaque1
through opaque17 (o1, o2, o5, o7, o9o11,
and o13o17), the semidominant floury mutants
(fl1, fl2, and fl3), the dominant mutants Mucro-
nate (Mc), and Defective endosperm B30
(De-B30). The position of these mutations and
zein genes on maize genetic map is shown in
Fig. 4.2. The o2 and De-B30 mutants are
located on chromosome 7, fl2 on chromosome
4, fl3 on chromosome 8, and o7 on chromosome
10. Chromosome 2 carries o4, o8, fl1, and
Mc mutants.
It has been reported that recessive mutations
affect regulatory genes, whereas semidominant
and dominant mutations affect the storage
proteins. All these mutations are associated
with increase in non-zein protein content and
decrease in the level of zein subunits. However,
premature death of the mutant seedlings and the
complex genetics behind lysine trait hinder
utilization of these mutant genes except for o2
in corn breeding and production (McWhirter
1971; Nelson 1979, 1981). Epistatic interactions
have also been reported among various regu-
latory mutants; o2 and o7 are epistatic over fl2,
whereas o2 and Mc have synergistic effect
(Prasanna and Sarkar 1991).
52
o13
z1C-1
z1A-1
z1D f12/ z1C-2
o8 cp2/o12
z1A-2
fl1/04
Mc
o1
1 2 3 4
Chromosome
Fig. 4.2 Map position of zein genes and mutants (Source: Gibbon and Larkin 2005)
4.4 Beginning of Genetic
Manipulation of Protein
Quality
The development of quality maize begins
initially with the manipulation of o2 and fl2
mutants singly or in combination. Later on, it
was realized that fl2 conversion programs were
difficult to handle as the kernel expression and
quality depended on dosage effect. On the other
hand, o2 mutants have simple recessive inheri-
tance, no dosage effect, and easily scorable phe-
notypic expression as opaque kernels help in
identification of the presence/absence of o2
gene. The intensive work for utilization of o2
mutation in breeding programs was continued
in 1969 at CIMMYT, Mexico. All potentially
important maize material was converted
to opaque2 with varying degrees of backcrosses.
India was among the first few countries in the
world to focus on improvement of maize quality
and a research program initiated in 1966 under
the All India Coordinated Maize Improvement
Project (AICMIP). Three o2 composites, namely,
Shakti, Rattan, and Protina, were developed
and commercially released in 1970 (Dhillon
and Prasanna 2001; Prasanna et al. 2001). The
o2 composites and hybrids were experimentally
tested and grown commercially in Brazil,
Colombia, India, the USA, South Africa,
Y. Vikal and J.S. Chawla
DeB30
o2
zp50/27
zp15
o5
dzs10
sh4-o9 pro1/o6
fl3
o15 o7
o14
5 6 7 8 9 10
Yugoslavia, and Hungary during the late 1960s
and early 1970s. The o2 maize had some pleio-
tropic effects such as reduced grain yield, chalky
and dull kernel appearance, susceptibility to ear
rots and stored grain pests, and slower drying of
grain following physiological maturity. Due to
these limitations, the o2 maize did not become
popular with farmers as well as with consumers,
despite its nutritional superiority. The kernel
became soft and more prone to mechanical dam-
age that decreased the yield by 815 %, and the
plants were more susceptible to fungi and insects
(Lambert et al. 1969; Salamini et al. 1970).
The search then continued by exploiting other
mutants in breeding programs such as o6, fl3,
and Mc, but none of them showed any additional
advantage over o2 and declined the efforts for
enhancing the nutritional value of maize kernel.
Double-mutant combination of o2su2 had vitre-
ous and small kernels and protein quality at par to
o2 but reduced grain yield by 1525 %, whereas
o2fl2 double-mutant combination resulted in
vitreous kernels only in few genetic back-
grounds. Despite the discouraging results,
researchers at CIMMYT, Mexico; the University
of Natal, South Africa; and Crows Hybrid seed
company at Milford, IL, continued their efforts
for improving the protein quality of maize. The
only viable strategy that came out was to
combine o2 with genetic modifiers of o2 for
rewarding results.
4 Molecular Interventions for Enhancing the Protein Quality of Maize
4.5 Development of Quality
Protein Maize (QPM)
Maize breeders began to identify modifier genes
(mo2) that alter the soft, starchy texture of the
endosperm, giving it a normal appearance while
maintaining the increased essential amino acid
content of o2 germplasm. During the process of
converting normal maize to o2 version, partially
hard endosperms were observed by Paez et al.
(1969). The plant breeders at CIMMYT, Mexico
(Vasal et al. 1980), and Pietermaritzburg, South
Africa (Gevers and Lake 1992), developed hard
endosperm o2 mutants, designated as quality
protein maize (QPM). QPM looks and performs
like normal maize, except that their nutritional
value gets improved. Since the mid-1990s, QPM
was tested at multiple research stations all over
the world, with encouraging results. Data from
32 locations across Africa, Asia, and Latin
America showed that QPM hybrids were capable
of outperforming the normal in some of the
poorest parts of countries such as China, Mexico,
Ghana, and Peru. In India, it led to the develop-
ment of a modified, nutritionally superior o2
composite in 1997, designated as Shakti-I.
The QPM research gained further momentum
by the launch of National Agricultural Tech-
nology Project (NATP) on QPM in 1998 by the
Indian Council of Agricultural Research (ICAR).
In this project, a multidisciplinary team of
multi-institutes involving Directorate Maize
Research (DMR), New Delhi; Punjab Agricul-
tural University (PAU), Ludhiana; Chaudhary
Charan Singh Haryana Agricultural University
(CCSHAU), Karnal Center; Acharya NG Ranga
Agricultural University (ANGRAU), Hyderabad;
and Rajendra Agricultural University (RAU),
Pusa, wherein the QPM germplasm received
from CIMMYT was acclimatized to suit the
local agroclimatic conditions in India. The lines
were evaluated for their productivity and
deployed in combination breeding, which led to
release of first QPM three-way cross hybrid,
Shaktiman-1, in 2001 followed by the release of
the first QPM single-cross hybrid, Shaktiman-2,
in 2004. Another QPM single-cross hybrid,
HQPM-1, is the first yellow grain QPM single-
53
cross hybrid released for its cultivation across the
country in 2005. Later in the series of QPM,
Shaktiman-3, Shaktiman-4, HQPM-5, HQPM-7,
and Vivek QPM-9 were released. Vivek QPM-9
has a unique distinction of the first molecular-
marker-assisted (MAS) converted product of
normal hybrid Vivek-9. The cultivation of QPM
hybrids will ensure higher income to farmers
as well as nutritionally superior food to the
consumers. Therefore, the QPM can be a strong
support to the mission of food and nutritional
security of the country particularly in underpriv-
ileged and tribal regions, where maize is con-
sumed as a staple food. QPM will also ensure
quality feed for poultry and animal sector, which
are the largest consumers of maize in India.
4.6 Molecular Basis of o2
and Modifier Gene Action
The breeding of QPM involves three genetic
systems: (i) the recessive mutant allele of the o2
gene, (ii) the endosperm hardness modifier genes,
and (iii) the amino acid modifier genes influencing
free amino acid content in the endosperm. Isola-
tion and characterization of o2 gene revealed that
it encodes a transcriptional factor that regulates
the expression of zein gene and a gene encoding a
ribosomal inactivating protein (Schmidt et al.
1990; Lohmer et al. 1991; Bass et al. 1992). The
o2 mutation typically reduces the level of 22-kDa
-zein and enhances the synthesis of a number of
non-zein proteins, particularly EF-1, that contain
relatively higher levels of lysine and tryptophan
(Damerval and Devienee 1993; Habben et al.
1993; Gaziola et al. 1999). In higher eukaryotes,
EF-1 is encoded by a multigene family. In maize,
there are 1015 genes, five of which are expressed
in endosperm (Carneiro et al. 1999). Wu et al.
(2002) identified two significant QTLs (on
chromosomes 2S and 4S) and two suggestive
QTLs (on chromosomes 5S and 6S) for EF-1.
The opaque2 gene is also known to be involved in
the synthesis of the enzyme lysine-ketoglutarate
reductase that is associated with free lysine degra-
dation. As a consequence, in the grain with o2
mutation, a dramatic reduction in this enzyme
leads to a corresponding increase in free lysine
54
Fig. 4.3 Kernels on a light
table showing varying
degrees of endosperm
modifications
in the endosperm (Brochetto-Braga et al. 1992).
The o2 allele is inherited in a simple recessive
manner, and the presence of o2 allele in homozy-
gous state (o2o2) is the prerequisite for the entire
process of obtaining high-lysine/high-tryptophan
maize.
The genetic modifiers are complex in their
inheritance and they are additive in nature. The
expression of modifiers varies with respect to
kernel modification, density, and biochemical
composition of the grain of the genetic back-
ground. Genetic analysis has shown that there
are two major modifier genes: one modifier
locus is tightly linked with -zein (27 KDa)
encoding sequence near the centromere of chro-
mosome 7, while the other is near the telomere of
chromosome 7L (Lopes et al. 1995). Using two
independently developed QPM lines, Holding
et al. (2008) mapped several major o2 modifiers
(Opm) QTLs to chromosomes 1, 7, and 9. In
addition, five other QTLs are also present: one
each on chromosomes 5S, 7L, and 10S and two
on chromosome 1L. So far, 10 gene modifiers
have been identified. A microarray hybridization
and real-time PCR performed with RNA from
true breeding o2 progeny with vitreous and
opaque kernel phenotypes identified several can-
didate genes that were upregulated and mapped
at or near the Opm QTLs, suggesting linkage of
o2 endosperm modification with these differen-
tially expressed genes. One of the QTLs is linked
to the 27-kDa -zein locus on chromosome 7S.
Moreover, QPM lines have 23-fold higher
levels of the 27-kDa -zein, but the physiological
significance of this increase is not known.
Because the genes encoding 27- and 16-kDa -
zein are highly conserved in DNA sequence,
Wu et al. (2010) introduced a dominant RNAi
transgene into a QPM line (CM105Mo2) to get
rid of their expression. Elimination of -zeins
Y. Vikal and J.S. Chawla
disrupts endosperm modification by o2
modifiers, signifying their hypostatic action to
-zeins. Abnormalities in protein body structure
and their interaction with starch granules in the
F1 with Mo2/+; o2/o2; RNAi/+ genotype sug-
gest that -zeins are essential for restoring
protein body density and starch grain interaction
in QPM. RNAi is to eliminate pleiotropic
effects caused by o2, resulting in protein bodies
formation such as honeycomb-like structures that
clearly demonstrate that -zeins play a mechanis-
tic role in QPM molecular breeding. Proteomic
analysis revealed that altered starch structure was
associated with endosperm modification in QPM
as granule-bound starch synthase I (GBSS I)
level was increased in modified kernels, which
alters the amylopectin branching in QPM.
Compared with normal and soft o2 genotypes,
amylopectin in QPM had reduced levels of inter-
mediate length -1, 4-linked glucose chains. As a
consequence, the QPM starch swelled more than
the normal and formed tight contacts between
starch granules in mature endosperm, resulting
in vitreous kernel phenotype. However, the
mechanism by which modifier genes create a
vitreous phenotype is yet to be understood. The
beneficial alleles of the modifying loci that con-
trol -zein production can be selected using a
rapid, low-cost, light table method. Due to segre-
gation of genes for endosperm hardness (or soft-
ness), varying degrees of softness/hardness are
expressed in the endosperm of segregating
generations (i.e., varying levels of opaqueness
are observed on a light table (Fig. 4.3). Gradation
in the opaqueness is scored on a 15 scale for
easy description of the various classes. Score 1 is
given for fully modified kernels (0 % opaque-
ness), scores 2, 3, and 4 for 25 %, 50 %, and 75 %
opaqueness, respectively, whereas score 5 for
soft, chalky, and 100 % opaque kernels.
4 Molecular Interventions for Enhancing the Protein Quality of Maize
The modified o2 version maize lines have
varying levels of lysine and tryptophan content.
This indicates presence of amino acid enhancer
genes (minor modifying genes), which neces-
sitate systematic biochemical evaluation of
lysine and tryptophan levels in each segregating
generation. At least three genes associated with
lysine level have been mapped to locations
on chromosomes 2, 4, and 7 (Wang et al. 2001).
Ten significant and one suggestive QTL for free
amino acid (FAA) content were identified on
all 10 chromosomes (Wu et al. 2002). The QTL
on chromosome 2L is coincident with genes
encoding two aspartate kinase enzymes, which
control important steps in amino acid biosynthe-
sis and lysine degradation pathways. A negative
correlation was observed between endosperm
texture trait and amino acid content (Gutierrez-
Rojas et al. 2008). Therefore, their regular moni-
toring at each step is essential as one could
end up with a maize cultivar having the o2o2
genotype with lysine and tryptophan levels
equivalent to those in normal maize (Krivanek
et al. 2007).
4.7 Molecular Interventions for
Enhancing the Protein Quality
The expression of the o2 allele is specific to
seeds, and recessive, conventional introgression
approaches require the inclusion of a selfing
progeny test to monitor the introgression of
the o2 allele within each backcross (BC) popula-
tion, and also higher plant population, enormous
time duration, labor, and spatial resources are
required. Numerous advantages such as reduced
time and population size are known to accrue to
the breeder by use of marker-facilitated genotype
selection rather than classical phenotypic selection
or by genetic engineering for endosperm-specific
expression of high lysine and tryptophan content.
4.7.1 Molecular Breeding for QPM
Mapping and tagging of agriculturally important
genes have been greatly accelerated by an array
55
of molecular markers in crop plants. The use of
DNA markers in backcrossing has greatly
increased the efficiency of selection. The practi-
cal application of molecular markers in crop
improvement is in marker-assisted selection
(MAS), which is of great importance as it helps
in improving the efficiency of plant breeding
through specific transfer of genomic regions of
interest (foreground selection) and accelerating
the recovery of the recurrent parent genome
(background selection). MAS have been more
extensively employed for simply inherited traits.
In 2001, the primer sequence of three SSR
markers (phi112, umc1066, and phi057), all at
the o2 locus, were released at the website www.
agron.missouri.edu, which facilitated the study
and application of the o2 gene. Phi057 is located
between G box and 3 upstream ORF in the leader
sequence of the o2 gene, and its mutation
can affect transcription of the o2 gene. The
umc1066 and phi057 are located in exon 1 and
exon 6, respectively; these are the two largest
exons among the six exons within the o2 gene.
To follow MAS, there are three main steps:
1. Validation of markers, i.e., the markers for the
target gene should give polymorphism bet-
ween recipient and the donor parent.
2. Foreground selection of the target gene with
linked molecular markers and recombinant
selection for recurrent parent allele at markers
flanking the target allele. Flanking markers
are used to select rare individuals that are the
result of recombination near the target gene,
thus minimizing the effects of linkage drag
(Ibitoye et al. 2010). Using very close markers
is the only way to reduce linkage drag,
substantially.
3. Marker-assisted background selection for
noncarrier chromosomes. Using markers, it
can be achieved by BC2 generation (Visscher
et al. 1996; Hospital and Charcosset 1997,
Frisch et al. 1999a, b), thus saving four BC
generations accelerating the rate of recurrent
parent genome recovery.
The utility of three maize SSR markers pres-
ent within the o2 gene in molecular-marker-
assisted selection for o2 has been successfully
used for conversion of non-QPM to QPM lines
56
in many parts of the world including CIMMYT,
Mexico; VPKAS, Almora; and PAU, Ludhiana,
in India and elsewhere in Asia and Africa (Babu
et al. 2005; Manna et al. 2005; Jompuk et al.
2006; Gupta et al. 2009; Agrawal and Gupta
2010; Zhang et al. 2010). At PAU, Ludhiana,
three microsatellite primers, viz., phi112,
umc1066, and phi057, within the gene o2 have
been surveyed among non-QPM lines (LM5,
LM6, LM11, LM12, LM13, and LM14), CML
lines (161, 163, 170, 189, 193, and 165), DMR
lines (7, 11, 561, 564, 568, 72, and 74), and
derived QPM lines (in different generations,
S1S4) for their validation. With phi057, three
alleles were detected, and this primer clearly
distinguished between QPM and normal lines.
With primer phi112, null allele was observed in
most of the QPM lines except CML 189, DMR 7,
DMR 568, and few derived QPM lines. With
primer umc1066, only two of the non-QPM lines,
viz., LM11 and LM13, were distinguished from
QPM lines (unpublished data). Xu and Crouch
(2008) also reported that umc1066 is easily
visualized on agarose gels but is commonly not
polymorphic in CIMMYT breeding populations,
while phi112 is a dominant marker and hence
cannot be used in the identification of
heterozygotes in F2/BC populations. It is best to
use three markers together in MAS for high-
lysine maize materials (Babu et al. 2005; Danson
et al. 2006; Mboogoi et al. 2006). A rapid line
conversion strategy using MAS program is given
in Fig. 4.4. Using combination of MAS and phe-
notypic selection techniques, linkage drag can be
reduced by selecting for flanking markers of recip-
ient allele type (selection of either single or double
recombinants) and recovery of the maximum
amount of recurrent parent genome (9095 %)
coupled with higher lysine and tryptophan content
within a short span of 3 years, which would other-
wise require 10 generations by conventional
backcrossing method.
Danson et al. (2006) used a modifier marker,
umc 1216, to select modifiers for the o2 pheno-
type showing two peaks for the non-QPM at
112 bp and 115 bp, while only one peak for the
QPM donor at 115 bp. However, Micic-Ignjatovic
et al. (2008) reported this marker as
Y. Vikal and J.S. Chawla
monomorphic. So, this marker could not be
employed in MAS for selection of modified endo-
sperm. Effective markers associated with
modifying loci for both endosperm hardness and
amino acid levels need to be identified. Besides,
phenotypic selection is applied for grain hardness
and amino acid content; the rapid line conversion
strategy using MAS was followed to develop
QPM hybrid Vivek QPM 9 in India at VPKAS,
Almora (Gupta et al. 2009), which was released in
2008 At PAU, Ludhiana, non-QPM lines, viz.,
LM11, LM12, LM13, and LM14, have been
converted into QPM versions, which are parental
lines of two important hybrids, Buland (LM11 
LM12) and PMH1 (LM13  LM14).
Another high-lysine mutant opaque16 (o16)
has been isolated from the Robertson Corn
Mutant library in Guizhou Institute of Upland
Food Crops, China. It has been mapped on long
arm of chromosome 8 and umc1141 marker is
linked to this gene. Double-recessive mutant
effect of the o16 with o2 was evaluated on lysine
content and the double-recessive hybrid showed
6 % higher lysine content than both the parents
(Yang et al. 2005; Zhang et al. 2010). This infor-
mation may be useful for marker-assisted selec-
tion and gene pyramiding in high-lysine maize
breeding programs.
4.7.2 Transgenics for QPM
Efforts in genetic transformation are focused on
developing a dominant opaque2 trait in maize.
RNA interference (RNAi) has been used to spe-
cifically suppress -zein production in transgenic
corn, resulting in doubling of the lysine content of
corn grain. Segal et al. (2003) used RNAi technol-
ogy to engineer transgenic maize for 22-KD -
zein gene, producing a dominant opaque pheno-
type. The phenotype segregated in a normal Men-
delian fashion and eliminated 22-kD zein without
affecting the accumulation of other zein proteins.
It was also found that the dominant phenotypes
generated were correlated with increased lysine
content. Similarly in another experiment, 19-kD
alpha-zeins got reduced by using antisense trans-
formation constructs (Huang et al. 2004, 2005). In
4 Molecular Interventions for Enhancing the Protein Quality of Maize 57

Fig. 4.4 Marker-assisted foreground and background selection scheme for quality protein maize (Source:
Babu et al. 2005)

both the studies, the increase of lysine was far
below than is reported in o2o2 maize. Huang
et al. (2006) changed the gene construct and
used a double-stranded RNA (ds RNA) to sup-
press the 19-kD and 22-kD -zein gene families.
Thus they could achieve the lysine 5.62 % and
tryptophan 1.22 %, which is higher that is achiev-
able with opaque2. This approach of using ds
RNA looks promising. While the dominant nature
of the antisense transgene is a definite advantage
as compared to recessive allele of o2, the opaque
endosperm still needs to be modified by endo-
sperm modifier genes whose epistasis with the
transgene has not yet been tested.
Alternatively, the plant lysine metabolic
pathway provides possible enzyme targets for
genetic engineering to increase free lysine
content in corn grain. As shown in Fig. 4.5,
lysine, along with methionine, threonine, and
isoleucine, is derived from aspartate; dihydrodi-
picolinate synthase (DHDPS) catalyzes the first
committed step of lysine biosynthesis. A bifunc-
tional enzyme, lysine-ketoglutarate reductase/
saccharopine dehydrogenase (LKR/SDH), is
responsible for lysine catabolism (Azevedo
et al. 2006; Stepansky et al. 2006).
The free lysine level in plant cells is thought
to be regulated by lysine feedback inhibition
of DHDPS and feed-forward activation of
LKR/SDH. Indeed, the expression of a lysine
feedback-insensitive DHDPS from Corynebacte-
rium glutamicum, CordapA, as well as the
58 Y. Vikal and J.S. Chawla

a b
Lysine metabolic pathway A bifunctional transgene
Asp to up-regulate lysine

1. Expression of a bacterial lysine feedback

insensitive DHDPS, CordapA, increases
Thr
lysine synthesis.
Met ASA
Ile DHDPS

Promoter intron CordapA 3UTR

Lys
KG

Srp LKR/SDH
2. An intron-embedded inverted repeat
Glu targeting LKR/SDH for suppression,
reduces lysine degradation.
AASA

Fig. 4.5 (a) The lysine metabolic pathway in plants.
In the biosynthesis of aspartate (Asp)-derived amino
acids, dihydrodipicolinate synthase (DHDPS) catalyzes
the committed step to (Lys) production and is sensitive
to feedback inhibition by Lys (red line). The bifunctional
lysine catabolic enzyme lysine-ketoglutarate reductase/
saccharopine dehydrogenase (LKRSDH) is activated by
excess lysine (green line). Other amino acids (shaded
box) and metabolites (open box) shown in the pathway
are threonine (Thr), methionine (Met), isoleucine (Ile),
glutamate (Glu), 3 aspartate semialdehyde (ASA), and
-aminoadipic-8-semialdehyde (AASA). (b) The design
of a bifunctional transgene cassette that simultaneously
deregulates lysine biosynthesis and reduces lysine degra-
dation (Source: Huang et al. 2008)

suppression of LKR/SDH have resulted in trans- biosynthesis and catabolism. GM crops with
genic corn with higher levels of free lysine higher levels of other important amino acids,
(Huang et al. 2005; Houmard et al. 2007). To namely, methionine, tryptophan, and threonine,
overcome lysine catabolism in the endosperm, are also expected. The opportunities for and the
maize plants were transformed with a single impacts of GM crops with enhanced nutritional
endosperm-specific bifunctional expression/ quality depend on public acceptance.
silencing transgene, which encodes a bacterial
feedback-insensitive DHDPS with a LKR/SDH
RNAi sequence in an intron (Frizzi et al. 2008).
4.8 Nutritional Superiority and
The suppression of LKR/SDH in the endos-
Biological Value of QPM
perm tissue increases free lysine to 1,324 ppm
(~30-fold increase). The combination of
Maize, being a potential crop in India, occupies an
CordapA expression and LKR/SDH suppression
important place as a source of human food (25 %),
in a single transgene produces over 4,000 ppm
animal feed (12 %), poultry feed (49 %), industrial
free lysine (~100-fold increase), the highest ever
products mainly as starch (12 %), and 1 % each in
reported in corn kernels. This construct resulted
brewery and seed. The maize grain accounts for
in a significant elevation in the seed lysine level,
about 1556 % of the total daily calories in diets of
proving that maize endosperm possesses enzy-
people in about 25 developing countries, particu-
matic activity responsible for both lysine
larly in Africa and Latin America, where animal
4 Molecular Interventions for Enhancing the Protein Quality of Maize
protein is scarce and expensive and consequently
unavailable to a vast sector of the population
(Prasanna et al. 2001). Research suggests that
QPM can help reduce protein deficiencies, partic-
ularly in young children, where maize dominates
in the diets (Vasal 2000). In QPM varieties,
leucine/isoleucine ratio was improved and became
better balanced which in turn is considered benefi-
cial as it helps to liberate more tryptophan for more
niacin biosynthesis, thus helping to combat pella-
gra (Prasanna et al. 2001). The other nutritional
benefits of QPM include higher calcium and car-
bohydrate (Bressani 1995) and carotene utilization
(De Bosque et al. 1988).
Because of the 60100 % increase in these
two essential amino acids, increased digestibil-
ity, and increased nitrogen uptake relative to
normal-endosperm maize, the biological value
of QPM is about 80 %, whereas that of normal
maize is 4057 % (Bressani 1992). The lysine
content of o2 maize is 2.54.0 g/100 g of protein,
which is more than twice that of an endosperm
from the normal maize (1.3 g lysine/100 g pro-
tein). Only 37 % of common maize protein intake
is utilized compared to 74 % of the same amount
of o2 maize protein. A minimum daily intake of
approximately 125 g of o2 maize might guaran-
tee nitrogen equilibrium. This could not be
obtained by using even twice the amount of nor-
mal maize. The nitrogen balance index for skim
milk and o2 maize protein is 0.80 and 0.72,
respectively, which indicates that the protein
quality of QPM is 90 % of that of milk. Besides,
around 24 g of normal maize per kg of body
weight is required for nitrogen equilibrium, com-
pared to only around 8 g for QPM (Bressani
1995; Graham et al. 1980). This nutritional
value of the QPM genotypes meets the
requirements of preschool children for their pro-
tein needs (Mertz et al. 1964; Vasal 2000).
The decreased level of zein (527 %) in o2
maize in contrast to normal endosperm
(5459 %) considerably improves its nutritional
quality. Another important application of QPM is
as animal feed, especially for monogastric
animals such as pigs and poultry, as it results in
improved growth. Small-holding farmers (who
typically cannot afford balanced feeds) and
59
commercial farmers find this extremely remuner-
ative. Therefore, the development and adoption
of improved QPM cultivars have significant
potential to alleviate hunger, increase incomes,
and improve livelihoods.
4.9 Conclusion
Malnutrition is one of the important issues in
India and the world. Therefore, development of
QPM hybrids, which are widely grown in spe-
cific locations will provide food and nutritional
security and thereby help in reducing poverty.
QPM is likely to gain wider acceptance if QTLs
for kernel modification and enhancers for amino
acids are fine mapped to develop markers to
follow MAS for vitreous kernels and high levels
of lysine. It also provides an ideal platform for
stacking of other nutritional genes for enhanced
Fe and Zn content and low phytate content for
multiple benefits. However, the major constraint
in adoption of QPM hybrids is contamination
with normal maize pollen in field, resulting in
erosion of the trait in farmer-saved seed system.
It is essential to give training on good seed
production practices to the local communities
and development of linkage between the
seed producers, farmers, and the industry for
sustainable higher nutritional benefits of QPM
in the long term.
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 Part III
Nutritional Quality of Maize
 Maize: Grain Structure, Composition,
Milling, and Starch Characteristics 5
Narpinder Singh, Amritpal Kaur, and Khetan Shevkani

Abstract
Maize is the third important food grain after wheat and rice, and its
demand is increasing because of its increased use for biofuel production.
Starch is the main component of maize, which is produced by wet milling
process. Maize starch functionality varies with the starch structure and
composition, which vary with genotypes and cultural practices. The
average size of maize starch granules ranges between 1 and 7 m and
15 and 20 m, respectively, for small- and large-sized granules. Maize
starches exhibit a typical A-type pattern, in which double helices com-
prising the crystallites are densely packed. Sugary maize starch has lower
crystallinity, while waxy maize starch has greater crystallinity as com-
pared to normal maize starches. The sugary maize starch has lower
gelatinization temperature and enthalpy. The maize starch with long-
branch chain length amylopectin and higher crystallinity has higher gela-
tinization temperature and enthalpy. The maize products (canned, frozen,
and boiled sweet maize) have lower glycemic index (GI) than white rice
and wheat flour bread. Waxy maize starch is more rapidly digested and
have high GI than high-amylose starches. Thermal treatments such as
autoclaving, baking, steam cooking, and parboiling processes affect starch
digestibility and consequently the GI of maize-based products. Maize
also contains various bioactive constituents, such as carotenoids,
anthocyanins, and phenolic compounds, which vary with maize type.
Maize has a higher antioxidant capacity compared to wheat, oat, and rice.

5.1 Introduction

The global maize (corn) production during 2010

N. Singh (*)  A. Kaur  K. Shevkani
Department of Food, Science and Technology, Guru
Nanak Dev University, Amritsar 143005, India
e-mail: narpinders@yahoo.com
was 844 million tons (mt) from an area of about
162 million hectares (mha). The USA, China,
Brazil, Mexico, Argentina, Indonesia, India,
France, South Africa, and Ukraine are the top ten

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 65
DOI 10.1007/978-81-322-1623-0_5, # Springer India 2014
66 N. Singh et al.

Table 5.1 Production of maize during 20002010a (metric tons)

Country 2010 2005 2000 1995 1990
USA 316,165,000 282,261,000 251,852,000 187,969,000 201,532,000
China 177,540,788 139,498,473 106,178,315 112,361,571 97,213,875
Brazil 56,060,400 35,113,300 31,879,400 36,267,000 21,347,800
Mexico 23,301,900 19,338,700 17,556,900 18,352,900 14,635,400
Argentina 22,676,900 20,482,600 16,780,700 11,404,000 5,400,000
Indonesia 18,364,400 12,523,900 12,043,200 8,245,900 6,734,030
India 14,060,000 14,709,900 12,043,200 9,534,000 8,961,700
France 13,975,000 13,687,700 16,018,400 12,739,600 9,400,500
South Africa 12,815,000 11,715,900 11,431,200 4,866,000 9,180,000
Ukraine 11,953,000 7,166,600 3,848,100 3,391,800
Source: aFAOSTAT (2012)

maize-producing countries in the world
(FAOSTAT 2012). The USA is the largest pro-
ducer and consumer of maize in the world,
contributing around 316 mt to the total global
maize production during 2010 (FAOSTAT
2012). China is the second largest producer of
maize with a production of 177 mt. The maize
production in Brazil and India was 56 and
21.72 mt, respectively, in 2010. The maize pro-
duction of major maize-producing countries is
shown in Table 5.1. The production of maize in
the USA, China, Brazil, and India was merely 201,
97, 21, and 8.9 mt, respectively, in 1995. Japan,
Republic of Korea, Mexico, China, Spain, and
Egypt are the main maize-importing countries.
Japan imported 3.7 mt of maize during 2009. The
USA, France, Argentina, Brazil, Ukraine, and
Hungary are the main maize-exporting countries.
The USA, Argentina, and Brazil exported 47.8,
8.5, and 7.7 mt, respectively, of maize during
2009. Different maize classes and cultivars are
grown in different countries. There are five general
classes of maize flint, pop, flour, dent, and sweet
corn which differ significantly in physicochemi-
cal characteristics and end-use quality. The
variations in maize characteristics and end-use
quality are attributed by hereditary and environ-
mental factors. On the basis of starch composition,
maize is categorized into three classes: (1) waxy
maize, which contains almost 100 % amylopectin;
(2) high-amylose maize, with starch containing
amylose content between 40 and 70 %; and (3)
sugary maize, with lower starch content and higher
level of sucrose (Nelson and Pan 1995). The dull
(du) and sugary (su2) genes have been associated
with higher amylose content (Inouchi et al. 1984;
Boyer and Liu 1985). The color of maize ranges
from white to yellow, red, and purple. Blue-,
purple-, and red-pigmented maize kernels are
rich in anthocyanins with well-established antiox-
idant and bioactive properties (Adom and Liu
2002). The pigmented maize is rich in antho-
cyanins, carotenoids, and phenolic compounds,
which have antioxidant and bioactive properties.
In India, maize has become the third important
food grain after wheat and rice. The demand for
maize is growing throughout the world because
of its increased demand for biofuel production.
The biofuel production is also being supported
by government policies in many countries. This
will further contribute to the increasing demand of
maize in the future. This may cause an increase in
the price of maize in the international market.
Maize is dry milled into grit used in the produc-
tion of breakfast cereals, snacks, etc. Wet milling
of maize is done to produce starch, which is the
raw material for the production of dextrins, glu-
cose, fructose syrups, sorbitol, etc. The present
chapter focuses on the grain structure, composi-
tion, milling, and starch characteristics of maize.
5.2 Grain Structure
and Composition
Maize grain is composed of endosperm
(8283 %), germ (1011 %), pericarp (56 %),
and tip cap (0.81.0 %). The pericarp is the
5 Maize: Grain Structure, Composition, Milling, and Starch Characteristics 67

Table 5.2 Total phenolics, anthocyanin, carotenoid, and ferulic acid contents of different maize types

Carotenoids Ferulic acid

Total phenolicsa Anthocyanin contentb (mg/100 g
Maize types (mg/100 g of dry wt) (mg/100 g of dry wt) Lutein -carotene dry wt)
Whitec 260.7  6.1 1.33  0.02 5.73  0.18 4.92  0.18 120
Yellowc 285.8  14.0 0.57  0.01 406.2  4.9 33.6  1.2 102
Yellowd 551  3.8 70.2  0.9 140
Redc 243.8  4.6 9.75  0.44 121.7  12.1 20.2  1.9 130
Redd 465  4.4 85.2  2.2 153
Bluec 266.2  0.7 36.87  0.71 5.17  0.49 23.1  2.1 130
Blued 343  8.6 99.5  1.8 152
Oranged 215  5.1 30.6  0.9 164
Purpled 465  9.8 93.2  1.1 154
Blackd 457  7.4 76.2  2.2 151
High carotenoidc 320.1  7.6 4.63  0.06 245.6  9.4 45.8  3.9 153
a
Expressed as gallic acid
b
Expressed as cyanidin-3-glucoside
c
de la Parra et al. (2007)
d
Lopez-Martinez et al. (2009)

outermost layer that is characterized by high crude
fiber content, mainly consisting of hemicelluloses,
cellulose, and lignin. Hemicellulose is present in
the highest concentration in the crude fiber. Peri-
carp thickness varies in different maize types and
extends to the base of the kernel joining the tip
cap. The pericarp and tip cap constitute a
small portion of total kernel lipids. The endo-
sperm is composed of a large number of cells,
each packed with starch granules embedded in a
continuous matrix of protein. The cell wall
consists of non-starchy polysaccharides (-glucan
and arabinoxylan), proteins, and phenolic acids.
Maize grain has two types of endosperm floury
and horny. Floury endosperm contains loosely
packed starch granules surrounding the central
fissure, while horny endosperm has tightly
packed, smaller starch granules toward the periph-
ery. The grains with higher proportion of harder
endosperm are preferred for the dry milling. The
storage proteins of endosperm are located within
subcellular bodies, simply known as protein bod-
ies, and comprise the protein matrix. Protein bod-
ies are composed almost entirely of a prolamine-
rich protein fraction known as zein, which is
extremely low in lysine. Whole maize grain has
protein, starch, and fat content between 8 and
11.5, 68 and 74, and 4.0 and 5.5 %, respectively.
The endosperm contains around 85 % starch and
8.5 % protein content. Fat content of the endo-
sperm is relatively low (1 %). The lipids present in
endosperm contain more saturated fatty acids
compared to germ lipids. The germ portion is
characterized by a high fat (33 %) and protein
(18 %) content. Germ has low starch content
(8 %), and its oil is high in polyunsaturated fatty
acids. Germ oil is relatively stable due to the
presence of high levels of natural antioxidants
and considered good for health because of its
fatty acid composition, mainly consisting of
oleic and linoleic acids.
Maize contains various bioactive constituents,
such as carotenoids, anthocyanins, and phenolic
compounds that have many health-promoting
and disease-preventing properties. Maize has a
higher antioxidant capacity compared to wheat,
oat, and rice (Adom et al. 2005). Lutein and
zeaxanthin are main carotenoids in maize, while
- and -cryptoxanthin as well as - and -caro-
tene are present to lesser extent. These bioactive
compounds vary in different maize types (Singh
et al. 2011). Total phenolic and anthocyanin
contents in maize vary with the color (Table 5.2).
The anthocyanins present in blue maize come
from cyanidin and malvidin (mainly from
derivatives of the former), whereas those in red
maize come from pelargonidin, cyanidin, and
malvidin. Yellow maize has more carotenoids
68
than floury maize. Blue and white maize are low
in lutein and zeaxanthin content, whereas yellow
and high-carotenoid maize have a higher content
(de la Parra et al. 2007). Bioactivities of purple-,
blue-, and red-pigmented maize are associated
with the presence of anthocyanins. Purple maize
is an important source of anthocyanins with
potential application as natural food colorants
and antioxidants (Aoki et al. 2002). High-
carotenoid genotypes have the best overall phy-
tochemical profile, followed by yellow, blue, and
red maize. White maize genotypes have lower
antioxidant activity due to lower amounts of
anthocyanins and carotenoids. Maize is also
known to contain a wide range of phenolic
acids. Ferulic acid is an important phytochemical
in maize, and its concentration vary in different
maize types. The high-carotenoid maize contains
higher amount of total ferulic acid compared to
white, yellow, red, and blue maize. Most of the
ferulic acid in maize is present in bound form
(Adom and Liu 2002). The bulk of phenolics
(phenolic acids, flavonoids, and conjugated
amines) are concentrated in the pericarp and
aleurone layers as well as the germ, with traces
in the endosperm (Sen et al. 1994). The highest
concentration of anthocyanin pigments in maize
is present in the pericarp portion, whereas the
aleurone layer contains small amounts (Moreno
et al. 2005).
5.3 Milling of Maize
5.3.1 Dry Milling
Maize is dry milled to grit, meal, and flour, which
are used in preparation of flakes, extruded
snacks, breakfast cereals, tortillas, etc. In India,
traditionally, the maize is dry milled in
mechanized stone mills to get whole meal or
atta that is used in the preparation of unleav-
ened flat bread or roti. The objective of dry
milling of maize into grit is to get maximum
recovery of grit with minimum contamination
of hull, germ, and tip cap. Flow diagram of dry
milling process of maize is illustrated in Fig. 5.1.
The grains are cleaned to remove impurities,
N. Singh et al.
conditioned with cold/hot water/steamed, and
then tempered for varying time to equilibrate
the moisture. The conditioning makes the hull
and germ tough and endosperm mellow.
The grains are milled by either Beall de-
germinator or fluted roller mills to separate
germ and hull. The Beall de-germinator consists
of a cast iron cone rotating within a perforated
conical housing with protrusions on the inner
surface. These protrusions rub off the germ and
hull. The roller mills for de-germination consist
of a set of fluted rolls that rotate at differential
speeds (1.25:11.5:1) in the opposite direction.
The gap between the rolls is adjustable
depending upon the size of grain and grit. The
de-germination using roller mill is comparatively
simple; however, the separation of germ and hull
is relatively inefficient. After de-germination, the
material is passed through 35 pairs of rolls and
graded on purifier to get fractions of different
particle sizes, ranging from large hominy grits
to fine flour. The products obtained from dry
milling include flaking grits; coarse, medium,
and fine grits; coarse or granulated meal; and
fine meal. The separated germ is used for oil
extraction either by solvent extraction or screw
pressing.
5.3.2 Wet Milling
Throughout the world, large quantities of maize
are wet milled to produce starch and other valu-
able by-products such as gluten, germ, and bran.
Flow diagram of wet milling process of maize
is illustrated in Fig. 5.2. Wet milling of maize
involves steeping of maize in water for 3040 h
at 50  C in the presence of SO2 (0.02 %) to soften
the kernels. SO2 prevents fermentation and
facilitates the separation of starch from protein.
After steeping, the steep water is drained, and
grains are coarsely ground in Fuss mill to free the
germ from endosperm and hull. The Fuss mill
consists of two upright, toothed metal plates
within a bronze-lined chamber. One plate of the
mill rotates while other remains stationary. The
soaked grains are fed between these plates which
cracks open the grains and free the germ. The
5 Maize: Grain Structure, Composition, Milling, and Starch Characteristics
Fig. 5.1 Flow diagram
of dry milling process
of maize
germ portion is then separated using flotation
tanks, washed, dried, and expelled for oil extrac-
tion. The de-germed suspension consisted of
fiber, starch, and protein is then fine milled in
attrition or impact mill. Fiber is separated by
screening and centrifugation. The fiber is washed
and dewatered by pressing and finally dried. The
suspension consisted of starch and protein is sent
to batteries of hydro-cyclones where they are
separated on the basis of differences in their
densities. Both the starch and protein (gluten)
are dried separately. The starch is utilized for
production of a number of value-added products,
such as dextrose, fructose syrups, dextrins,
69
modified starches, and sorbitol, by chemical
and/or enzymatic processes. The gluten is dried
and commonly used as animal feed.
5.4 Starch Characteristics
5.4.1 Granule Morphology
Morphological characteristics of starches from
different maize types vary with the genotype and
cultural practices. The morphology of starch
granules depends on the biochemistry of the chlo-
roplast or amyloplast as well as physiology of the
70
Fig. 5.2 Flow diagram of
wet milling process of
maize
plant (Badenhuizen 1969). Scanning electron
micrographs of the starch granules from different
maize types are illustrated in Fig. 5.3. The average
size of small- and large-sized granules ranges
between 1 and 7 m and 15 and 20 m, respec-
tively. Normal maize and waxy maize starch
granules displayed spherical or angular shape,
while sugary maize starch displayed irregular-
shaped granules consisting of lobes (Sandhu
et al. 2007). Normal and waxy maize starch
granules had the smooth surface, whereas sugary
N. Singh et al.
maize starch granules displayed rough surface
(Perera et al. 2001; Sandhu et al. 2007).
5.4.2 Structure
Maize starch, like starches from other cereals,
consists of amylose and amylopectin. Amylose
is essentially a linear polymer of glucose, while
amylopectin is branched. Normal starch contains
25 % amylose and 75 % amylopectin. Maize with
5 Maize: Grain Structure, Composition, Milling, and Starch Characteristics
Fig. 5.3 Scanning electron micrograms (SEM) of starch granules from different types of maize (Source:
Sandhu et al. 2007)
amylose as high as 85 % (amylomaize) is also
reported. Waxy maize starch mainly consists of
amylopectin (99 %) and contains negligible amy-
lose (<1 %). Amylose and amylopectin packing
in the granules vary among the starches from
different species. The branches of the amylopec-
tin molecules form double helices that are
arranged in crystalline regions. X-ray diffractom-
etry is used to reveal the presence and
characteristics of the crystalline structure of the
starch granules. The A-, B-, and C-type
patterns are different polymeric forms of the
starch determined by X-ray diffractogram.
These types differ in the packing of the amylo-
pectin double helices. Maize and other cereal
starches exhibit a typical A-type pattern, in
which double helices comprising the crystallites
71
are densely packed with low water content.
Tuber starches show the B-type pattern, in
which crystallites are less densely packed and
have more open structure containing a hydrated
helical core (Tester et al. 2004). Sugary maize
starch showed a weak A-type pattern with lower
crystallinity as compared to other maize starches.
Sugary maize starch showed larger amount of
amyloselipid complex as compared to normal
and waxy starch (Singh et al. 2006). Waxy maize
starches were reported to have greater crystallin-
ity. Among the waxy maize starches, differences
in crystallinity have been observed and were
related to the amount of longer amylopectin
chains. Waxy mutant starch has been reported
to be more crystalline than regular cereal starches
(Singh et al. 2003; Vandeputte et al. 2003).
72
Fig. 5.4 Typical pasting/
RVA profile of maize
starch
5.4.3 Pasting Properties
The pasting profile represents change in the viscos-
ity of starch suspension during heating and cooling
under constant stirring conditions. The Brabender
Viscoamylograph and Rapid Visco Analyser
(RVA) are extensively used for measuring pasting
profile of different starches. The typical pasting
curve of maize starch is shown in Fig. 5.4. The
pasting curve shows a steep increase at a certain
point (pasting temperature), representing minimum
temperature required for initiation of gelatinization.
Waxy starches have lower pasting temperature than
the normal starches (Liu et al. 1997; Sandhu et al.
2007). The peak viscosity reflects the point of max-
imum swelling of the granules. The starches from
normal and waxy maize show sharp peak, whereas
that of sugary maize show the flatter peak (Li and
Corke 1999; Perera et al. 2001; Sandhu et al. 2007).
Waxy starches show higher peak viscosity than
normal and sugary starches. The higher peak
viscosity of waxy starches is attributed to the
absence of amyloselipid complex. The lipids
and phospholipids have been reported to form com-
plex with the amylose and long-branch chains of
amylopectin in the cereal starches, consequently
N. Singh et al.
severely retarding swelling and inhibiting amylose
leaching (Larson 1980). Waxy cereal starches
are reported to have negligible lipid contents,
swell rapidly, and achieve higher peak viscosity
(McPherson and Jane 1999). Normal starches
have much higher amylose content, which restricts
swelling and limits the increase in viscosity by
developing aggregated structure (Becker et al.
1981). Further heating of starch suspensions at ele-
vated temperatures resulted into breakdown in vis-
cosity. The breakdown viscosity reflects the shear
thinning of the starch gels. Waxy starches showed
greater breakdown and higher shear thinning of the
gels due to the absence of amylose (Bahnassey and
Breene 1994). Low viscosity and almost negligible
breakdown for sugary maize have been observed by
Li and Corke (1999), and Singh et al. (2006). Sug-
ary maize starches having less breakdown may be
due to the loosely branched amylopectin structure
(Campbell et al. 1994). Upon cooling, the
gelatinized starch molecules partially reassociate
to form gel (retrogradation), and this leads to an
increase in the viscosity. The rise in viscosity during
cooling varies in different maize starches, and
these differences are attributed to the variation
in amylose.
5 Maize: Grain Structure, Composition, Milling, and Starch Characteristics
5.4.4 Thermal Properties
Starch undergoes irreversible changes
characterized as gelatinization when heated
with excess of water. Gelatinization is the disrup-
tion of molecular order within the starch granule.
It results in granular swelling, crystallite melting,
loss of birefringence, viscosity development, and
solubilization. Differential scanning calorimeter
(DSC) has been widely employed to study starch
gelatinization. DSC endotherms provide infor-
mation on gelatinization behavior of starches as
onset temperature (To), peak temperature (Tp),
conclusion temperature (Tc), and enthalpy of
gelatinization (Hgel). To, Tp, and Tc of normal
maize starches range between 63.5  C and
68.4  C, 67.8  C and 71.6  C, and 73.2  C and
76.3  C, respectively, while the waxy maize
starches have a range of 64.469.1  C,
70.473.5  C, and 80.581.3  C, respectively
(Singh et al. 2006). Sugary maize starch shows
lower transition temperatures and Hgel
attributed to the presence of lower crystallinity.
Higher degree of crystallinity provides structural
stability to the granules and makes them more
resistant to gelatinization and consequently
results into higher transition temperatures
(Barichello et al. 1990). The starches with long-
branch chain length amylopectin show higher
gelatinization enthalpy, indicating that more
energy is required to gelatinize the crystallites
of long chain length. The difference in gelatini-
zation range T {2(TpTo)} among the starches
from different maize types also differ may be due
to the presence of crystalline regions of different
strengths in the granules (Banks and Greenwood
1975). The variability in T values in starches is
due to the difference in the degree of heteroge-
neous crystallites, which have slightly different
crystal strengths (Vasanthan and Bhatty 1996;
Gunaratne and Hoover 2002). The Hgel
followed the order: waxy maize starch > normal
maize starch > sugary maize starch (Perera et al.
2001; Sandhu et al. 2007). Higher Hgel for
waxy maize starches has been attributed to the
presence of greater amount of crystallinity than
that present in normal maize and sugary maize
73
starches (Singh et al. 2006). The high Hgel of
waxy maize starches suggested that the double
helices (formed by the outer branches of adjacent
amylopectin chains) that unravel and melt during
gelatinization are strongly associated within the
native granule. The Hgel for sugary maize
starch (su2) was reported to be lower as com-
pared to normal maize starch (Inouchi et al.
1984). Higher Hgel value of the waxy maize
as compared to the normal maize has been
observed by Liu et al. (1997). Perera et al.
(2001) reported the lower value of transition
temperatures of sugary maize as compared to
normal and waxy maize starches. The gelatiniza-
tion enthalpy values of starches has been
reported to be affected by factors such as granule
shape, percentage of large and small granules,
and the presence of phosphate esters (Stevens
and Elton 1971; Yuan et al. 1993). The lower
Hgel of baby maize starch may be attributed to
its small granule size and low amylose content.
5.4.5 Starch Digestibility
In humans, the starch is digested in the small
intestine. However, a part of it is not digested
in the small intestine and reaches the large intes-
tine where it is fermented by the gut microflora
that produces short chain fatty acids as end
products, which are known to promote the opti-
mal function of the viscera (Topping and Clifton
2001). This indigestible portion of starch is
known as resistant starch. During heating, starch
is gelatinized and the semicrystalline structure of
starch disintegrates. The gelatinized starch
tends to reassociate (recrystallization) to form
an ordered gel-like structure during cooling
known as retrogradation. Retrogradation process
during the storage of processed starchy foods
(noodles, bread, and cooked rice) affects the
digestibility and often consumer acceptability of
such foods. The retrogradation rate and its extent
in starch foods vary with starch properties
(molecular and crystalline structure) and storage
conditions (temperature, duration, water content,
etc.). Foods containing starches with higher
74 N. Singh et al.

Table 5.3 Glycemic index (GI) of maize, rice, and depending upon the rate of release and
wheat products absorption of glucose in the gastrointestinal
Food GI tract: rapidly digestible starch (RDS), slowly
Maize meal porridge 109 digestible starch (SDS), and resistant starch
Corn muffin, low amylose 102 (RS). RDS is the group of starches that can be
Corn flakes 84 rapidly hydrolyzed by digestive enzymes, SDS is
Maize chapatti 5964 the group that is digested at a relatively slow rate
Sweet corn, boiled 60 (Englyst et al. 1992), and RS is not digested by
Whole sweet corn (canned) 46 digestive enzymes and consequently is trans-
Whole sweet corn (frozen) 47
ferred into the colon. Waxy maize starch is
Corn tortilla (Mexican) 52
more rapidly digested than high-amylose starch,
Corn muffin, high amylose 49
attributed to more surface area per molecule of
Sweet corn on the cob (boiled, 20 min) 48
the amylopectin than amylose. Many health
White rice 64
Brown rice 55
benefits such as improved cholesterol metabo-
Parboiled rice 47 lism and reduced risk of type II diabetes and
Bulgur wheat 48 colon cancer have been associated with the con-
Puffed wheat 74 sumption of RS (Hoebler et al. 1999).
Wheat bread 70 Amylolysis susceptibility of normal maize starch
Wholemeal bread 69 is more, as compared to high-amylose maize
Source: Foster-Powell et al. (2002). Reference food is starch, may be due to the presence of surface
glucose pores and channels that facilitate enzymatic dif-
fusion (Zhang et al. 2006). The association
amylose content have higher retrogradation rates between amylose chains and their potential for
(Singh et al. 2006). The sugary maize starch with amyloselipid complex formation (Morita et al.
higher amylose has a higher retrogradation rate 2007), higher crystalline lamella thickness, and a
than normal maize starch (Singh et al. 2006). The thicker peripheral layer (Jenkins and Donald
retrogradation makes the starch resistant toward 1995) are the factors that make the high-amylose
breakdown by digestive enzymes and conse- maize starch granules resistant to amylolysis.
quently reduces the glycemic index (GI) Thermal treatments such as autoclaving, baking,
(Fredriksson et al. 2000). GI characterizes the steam cooking, and parboiling processes affect
carbohydrates consumed in the form of different the gelatinization and retrogradation processes
foods on the basis of the postprandial level of and, consequently, the formation of RS in
blood glucose (Jenkins 2007). Carbohydrates foods. Hi-maize is rich in amylose (80 %) and
that are quickly digested and release glucose RS (42 %), which is a commercial source of RS
rapidly into the bloodstream are considered to that is widely used in various baked goods. The
have a high GI. GI of maize, rice, and wheat chemical modifications of starches are carried
products is shown in Table 5.3. Canned, frozen, out to change the functional properties that also
and boiled sweet maize has lower GI (4647) as change the susceptibility toward the action
compared to white rice (64) and wheat bread of enzymes, e.g., esterification, etherification,
(70). Long-term intake of foods with a high GI and cross-linking of starch make it resistant to
is associated with obesity, diabetes, and cardio- -amylase and reduce digestibility (Hood and
vascular diseases. Foods with low GI are Arneson 1976).
recommended for the prevention of diseases
that are associated with metabolic disorders
Acknowledgment Financial assistance by the Depart-
(Englyst et al. 1999; Jenkins et al. 1981; Ludwig ment of Science and Technology, Govt. of India, to
2000). Starch is classified into three groups Prof. Narpinder Singh is acknowledged.
5 Maize: Grain Structure, Composition, Milling, and Starch Characteristics
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 Oil Improvement in Maize: Potential
and Prospects 6
Naveen Singh, Sujata Vasudev, D.K. Yadava, D.P. Chaudhary,
and K.V. Prabhu

Abstract
High-quality maize oil, having low level of saturated fatty acids, is highly
suitable for human consumption. It is considered to be better than most of
other edible oils due to its fatty acid composition and stability during
storage and cooking. There is about 34 % oil content in maize kernel.
However, more than 67 % oil is reported in high-oil corn genotypes.
High-oil lines, in general, have reduced yields. Large numbers of genes/
QTLs were reported to control this trait, thus, making it difficult to improve.
A combination of conventional breeding methods, marker-assisted selection
and transgenic approach would help in developing high-yielding genotypes
with enhanced oil content in maize.

6.1 Introduction feed, it has wide range of industrial applications

Maize is an annual crop with high productivity
and exceptional geographic adaptability that has
helped its cultivation to spread throughout the
world. It is one of the three most important food
grain crops in the world. In India, it occupies the
third position among food grains with respect to
production and productivity and ranks fifth for
the area under cultivation. It is used for human
food, livestock and poultry feed, industrial raw
material and as fodder. In addition to food and
N. Singh (*)  S. Vasudev  D.K. Yadava  K.V. Prabhu
Division of Genetics, Indian Agricultural Research
Institute, New Delhi, India
e-mail: n.singhk@rediffmail.com
D.P. Chaudhary
Directorate of Maize Research, New Delhi, India
from food processing to manufacturing of
ethanol.
Maize is one of the oldest human-
domesticated plants. Its origins are believed to
date back to at least 7,000 years ago when it was
grown in the form of a wild grass called teosinte
in Central Mexico. Recognizing its early poten-
tial as a major food crop, over time, the
Mesoamerican natives managed to improve the
crop, by systematically selecting certain varieties
for their desired traits. This process led to the
gradual transformation of teosinte to its present
day form known as maize, a name which is a
likely derivative of mahis, meaning source of
life for Taino people. Maize is also known as
corn in the United States, the worlds largest
producer, consumer and exporter of maize. This
reference to corn, rather than maize, in the
United States can be traced to the arrival of the

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 77
DOI 10.1007/978-81-322-1623-0_6, # Springer India 2014
78
early European settlers in the New World when
maize was referred to as Indian corn. Indian corn
is a type of maize known for its colourful kernel.
The term corn most likely originates from the
Germanic korn, which referred to any edible
grass. Maize occupies a dominant place in the
world economy and trade as an industrial grain
crop (White and Johnson 2003).
Plant breeding has been extremely successful
in improving the yield of maize. Advances made
by breeders in quality improvement resulted in
maize kernels with a wide range of structures
and compositions. By exploiting genetic variation,
the composition of the kernel was altered for both
the quantity and quality of starch, proteins and oil.
The great economical and nutritional value of the
maize kernel is mainly due to its high starch
content that represents 75 % of the mature seed
weight, along with the protein complement (10 %
of the mature seed weight), mainly found in the
form of zeins (storage proteins) and oil (4.6 %).
The intensive use of the maize kernel is due not
only to its high starch content but also to the oil
stored in the embryo. Oil, in fact, is the most
valuable coproduct from industrial processing of
maize grain, and it represents a source of high-
quality oil for humans (Hartings et al. 2012).
6.2 Status of Corn Oil in India
Cultivation of a large number of oilseed crops in
India ensures availability of a vast range of edi-
ble oils; therefore, edible purpose corn oil is not
very popular for domestic uses. In the United
States, corn oil constitutes 4.3 % of vegetable
oil production and about 7.2 % of domestic oil
utilization. In India, however, it is not an ingre-
dient in the traditional food recipes, and this is
the main reason for less popularity of corn oil for
cooking purpose. High price and lack of public
awareness of corn oil further restrict its populari-
zation in the country. Almost 2 mt of maize is
used in the starch industry, yielding about 70,000
tonnes of corn oil as a by product, annually. This
figure is expected to rise up to 8 mt by 2050,
which can produce about 0.3 mt tonnes of oil
N. Singh et al.
(Hegde 2012). Local factories are extracting
3.03.5 % oil from corn, which otherwise contain
at least 56 % of oil. Thus, in India, there is a
huge scope in increasing the production of corn
oil by improving the efficiency of oil extraction
from corn germs. Furthermore, efforts are needed
to develop maize hybrids with higher oil content
in the germ, which in turn will help in obtaining
higher oil yield from maize processing.
6.3 Quality of Corn Oil
Corn oil is valued for its high energy content.
Energy value of 100 g corn oil is 884 kcal as
against 86 kcal in case of equal amount of corn
meal. Good quality of cooking oil is usually
associated with elevated proportion of unsatu-
rated to saturated fatty acids. Maize oil is highly
regarded for its low levels of saturated fatty
acids, with an average 11 % palmitic acid and
2 % stearic acid, against relatively high levels of
polyunsaturated fatty acids such as linoleic acid
(24 %). Maize oil is comparatively stable since it
contains only small amounts of linolenic acid
(0.7 %) and high levels of natural antioxidants
(Val et al. 2009). Furthermore, presence of sig-
nificant amount of phytosterols (968 mg/100 g)
in corn oil improves its quality significantly by
reducing the amount of cholesterol absorption
from food. Corn oil also provides a good source
of vitamin E (21.11 mg ATE/100 g) and tocoph-
erol (14.3 mg/100 g). These are responsible for
increased shelf life of corn oil. Four isomers of
tocopherols are found in corn, viz., , , and ,
and out of these, -tocopherol is the most com-
mon. It has been found that genotype and envi-
ronmental factors affect fatty acid composition
of corn oil. The fatty acid composition of corn oil
is affected by position of kernels on the ear.
Palmitic and linoleic acid content of the oil
increases for kernels from the base to the tip of
the ear. On the other hand, oleic acid content of
the kernels decreases from base to tip. Sampling
of kernels in the central portion of the ear is
recommended for the analysis of fatty acid com-
position (Zheng et al. 2008).
6 Oil Improvement in Maize: Potential and Prospects
6.4 Importance and Merits
of Corn Oil
Most common use of corn oil is as salad oil,
cooking oil and in margarine. Preference of
corn oil over other vegetable oils is mainly due
to its stability during storage and cooking that
saves the consumers from the additional synthetic
antioxidants. The presence of tocopherols in corn
oil acts as natural antioxidants by protecting dou-
ble chemical bonds from oxidation. Tocopherols
along with low levels of linoleic acid (18:3) are
responsible for stability of corn oil. In addition, its
blend flavour and high smoke point (>200  C) are
mainly responsible for its popularity. Phytosterol
in corn oil is responsible for lower cholesterol
absorption in the body from cholesterol-
containing food materials, and it increases the
percentage of high-density lipoproteins (HDL) in
blood. Large numbers of cosmetic products prin-
cipally based on corn oil are available in the
markets. Crude corn oil can also be used as sur-
factant. There are reports in which corn oil has
been used to increase the efficiencies of fungal
biocontrol agents.
6.5 High-Oil Corn
It is estimated that 8084 % of the total oil is
located in the germ, 12 % in the aleurone layer
and 5 % in the endosperm. Normal corn is rich in
starch and provides around 4 % oil, whereas
high-oil corn (HOC) contains more than 6 % oil
(Lambert 2001). There is a 34 % increase of oil
in high-oil maize over normal corn. Additionally,
protein quality and quantity are increased to
some extent in high-oil corn due to larger germ
(including embryo and scutellum) size. High-oil
corn differs from normal corn with respect to
kernel anatomy and composition. The germ in
HOC has about 1 % higher total protein and it
also contains higher levels of essential amino
acids like lysine, threonine and methionine,
therefore, making it highly suitable for human
and animal consumption (White et al. 2007).
79
High-oil maize shows a greater feed efficiency
than normal maize in animal feed trials because
the oil imparts 2.25 times higher caloric content
than that of starch on dry weight basis and also
because of its fatty acid composition, mainly oleic
and linoleic acids (Gross and Kerr 1992). It has
been observed that when oil content is higher
(more than 6 %) in feed rations, the animals usu-
ally observe a significant feed response over nor-
mal rations. It has been reported from feeding
experiment that lambs on high-oil and high-
protein rations gained 7 % more weight, retained
more nitrogen and required 6 % less feed per
pound of grain than the normal diet (http://
expert.iasri.res.in/agridaksh/html_file/maize/Intro
duction_highoilcorn.htm). High-oil corn diets fed
to cows during the last 30 days of gestation
resulted in significantly greater body weight
gains and higher fat levels in the colostrum than
normal diets. Feeding trials on laying hens showed
that diet of 17 % protein with high-oil corn leads
to better egg to feed ratio than normal ration and
egg production and yield increased with the high-
protein high-oil diets. Skin and plasma pigmenta-
tion was also improved in chicks fed with high-oil
diets. Feeding trials with lactating dairy cattle
showed 12 % greater intake of dry matter for
high-oil diet. On an average, feeding trials
suggested an advantage of 7 % or more with
high-oil corn rations in chicken, swine, dairy cat-
tle and sheep over normal rations. High-oil corn,
therefore, plays an important role in feed rations
due to its nutritional advantage and greater energy
per unit of feed.
Protein concentration in high-oil hybrids is
higher and protein quality is enhanced because
of the larger scutellum size compared with normal
corn. Amino acid lysine increased from 0.244 %
(normal corn) to 0.297 % (high-oil corn). High-oil
corn hybrids usually have higher proportion of
yellow pigments (carotenoids, xanthophylls,
etc.), which are beneficial in poultry feed and
play important biological functions in mankind.
Improvement in the oil content of the hybrids
is most important for the growth of corn oil
industry and production of low-cost oil. This, in
turn, will help in acceptance of this oil by the
consumers.
80
6.6 Genetic Resources
Knowledge of genes involved in determining
quality-related traits (structure and chemical
diversity) of starch, proteins, oil and other
compounds is important for improving the indus-
trial, nutritional and food-making properties of
the maize grains. Corn kernel oil content ranges
between 1.2 and 20.0 on weight basis. In maize,
kernel oil content is considered a quantitative
trait controlled by many genes with small effects
(Dudley 1977). The number of genes controlling
this trait estimated by quantitative genetic and
biochemical methods was found to be up to 69
(Dudley and Lambert 1992). Genetic mapping of
oil traits indicates that multiple (>50) QTLs are
involved in lipid accumulation (Laurie et al.
2004), thus making it difficult to improve.
Oil and starch are accumulated in different
compartments of kernel. Eighty-five percent of
the oil is stored in the embryo, whereas 98 % of
the starch is located in the endosperm. Studies
on the biosynthesis of oil indicate that plant oil
is synthesized from glycerol-3-phosphate and
fatty acyl-CoA in the endoplasmic reticulum as
triacylglycerols (TAGsesters of fatty acids and
glycerol) (Baud and Lepiniec 2010, Barthole et al.
2012). Fatty acids are synthesized from acetyl-
CoA in plastid and then transported to the cyto-
plasm in the form of fatty acyl-CoA where they
are used for the acylation of the glycerol-3-phos-
phate. Resulting TAGs are stored in specialized
structures called oil bodies. The primary determi-
nant of lipid content in maize kernel is the genetic
makeup (Lambert and Hallauer 2001). In spite of
a good understanding of the oil biosynthetic path-
way in plants and identification of the genes
involved, the molecular basis for oil quantitative
trait loci (QTLs) is largely unknown. However,
Zheng et al. (2008) found an oil QTL (qHO6)
affecting maize seed oil and oleic acid content.
This QTL encodes an acyl-CoA:diacylglycerol
acyltransferase (DGAT1-2), which catalyses the
final step of oil synthesis. Evidence has been
obtained that genetic variation also exists for the
fatty acid composition of the kernel (Lambert
2001). Studies indicate that the inheritance of
N. Singh et al.
oleic, linoleic, palmitic and stearic acid content
is complex and under polygenic control. Molecu-
lar characterization of fatty acid desaturase-
2 (fad2) and fatty acid desaturase-6 (fad6) reveals
that fad2 and fad6 are not associated with QTLs
for the ratio of oleic/linoleic acid. This suggests
that some of the QTLs for the oleic/linoleic acid
ratio involve other genes that may influence the
flux via enzymes encoded by fad2 or fad6 (Motto
et al. 2011).
Application of new technologies, such as tran-
scription profiling, metabolic profiling and flux
analysis, should prove valuable to more precisely
identify the genes and enzymes, determining the
composition of maize oil (Hartings et al. 2012).
In addition, identification of transcription factors
or other regulatory proteins that control oil bio-
synthesis or embryo development will be partic-
ularly useful for biotechnological approaches in
the future. Arabidopsis orthologs in maize that
trigger genes involved in TAG metabolism
(LEAFY COTYLEDON1-2, i.e., ZmLEC1), and
those necessary to mediate the regulators towards
late glycolytic and oil metabolism of the tran-
scription factor WRINKLED1 (i.e. ZmWRI1a and
ZmWRI1b), have been identified (Pouvreau et al.
2011). In maize, both genes are preferentially
expressed in the embryo and exhibit a peak of
expression at kernel maturity. ZmWRI1a is
induced by ZmLEC1 (Shen et al. 2010). Further-
more, transcriptomic analysis of ZmWRI1a
overexpressing lines led to the identification of
putative target genes of ZmWRI1a involved in
late glycolysis and fatty acid or oil metabolism.
6.7 Breeding for Oil Yield
Efforts directed toward increasing oil content
of maize kernels through breeding have been
successful, but unfortunately high-oil lines have
significantly reduced yield (Moose et al. 2004)
as, compared to starch, synthesis of oil is an
energy-consuming process. In general, high-oil
corn hybrids yield 510 % lower than normal
corn; therefore, attempts are being made to
enhance oil content without compromising
yield potential. Continuous genetic selection
6 Oil Improvement in Maize: Potential and Prospects
has steadily improved agronomic performance
and yield of corn hybrids with higher oil content,
thus, narrowed the yield gap between high-oil
corn and normal corn hybrids. Alexender
(1988) started the breeding programme of high-
oil maize using recurrent selection with the
genetic diverse synthetic, and after 27 selection
cycles, the oil concentration of the improved
population reached 21.2 %. Song et al. (2004)
developed high-oil maize from synthetic variety
Zhongzong No. 2 through a succession of 18
recurrent selections and increased the oil content
from 4.71 % to 15.55 %.
The genotype of the sporophyte greatly
determines the amount of oil produced by high-oil
hybrids so that these hybrids may have a more
efficient energy-trapping system. It has been
observed that the female parent had the largest
influence on oil percentage of the kernel as com-
pared to the male parent. Even then, xenia effects
existing in corn for total oil content possibly
resulted from an increase in grain size and
increased oil content in the germ as high-oil content
of the pollen source increases the proportion of
germ in the kernel. Similar relationship exists for
oil per cent in the germ. Under open pollinated
conditions, the oil levels of the lower-oil hybrids
may be increased, and the oil levels of the higher-oil
hybrids may be reduced because of xenia effects.
Genotype  environment interactions and
crop management practices such as dosage of
fertilizers has been reported to influence oil con-
tent in high-oil genotypes. However, effect of
genotype on oil values seems to be greater. Vari-
ation in total oil content is not associated with
total protein content but is positively correlated
with per cent germ protein and relative concen-
tration of tryptophan in the kernel. Association of
increased oil content with reduced ear length,
smaller ear diameter, lighter kernel weight,
reduced plant height and ear height and early
flowering has been reported. Successful breeding
for high oil content in the Illinois high-oil strains
has mainly been achieved through an increase in
embryo size (Moose et al. 2004). To enhance the
oil content in maize kernels, the relative propor-
tion of the oil-rich embryonic tissue within the
grain needs to be increased. However, the
81
molecular mechanism responsible for this has
not been understood well.
Recombinant DNA technology is being utilized
to enhance the quality and quantity of maize oil. In
the transgenic maize lines, expressing the wheat
Purindoline a and b (PINA and PINB) genes, the
total oil content of the kernel was increased by
25 % (Zhang et al. 2010). An examination of
carbon metabolism in maize embryos suggested
that the flux of carbon through NADP-ME may
constitute a metabolic bottleneck (Alonso et al.
2010). The oil content of the kernel is, thus, posi-
tively correlated with malic enzyme activities in
maturing embryos (Doehlert and Lambert 1991),
which makes NADP-ME an attractive target for
engineering high-oil trait in maize. Numerous bio-
technological approaches have been devised to
maximize the flow of C into oil by overexpression
of enzymes of the TAG assembling network in
oilseed crops. In maize, several attempts have
been made to overexpress diacylglycerol
acyltransferases (DGAT). The embryo-specific
overexpression both of maize DGAT12 and of
fungal DGAT2 (Zheng et al. 2008; Oakes et al.
2011) resulted in significant increase in kernel oil
content. These studies provide insights into the
molecular basis of natural variation of oil and
oleic acid contents in plants.
Seed-specific expression of ZmWRI1 enh-
anced oil accumulation in transgenic maize with-
out any associated abnormalities. It was also
found that ZmWRI1 not only increases the fatty
acid content of the mature maize grain but also of
certain amino acids (Lys, Glu, Phe, Ala and Val)
and of several compounds involved in amino
acid biosynthesis (Pouvreau et al. 2011). How-
ever, expression of ZmLEC1 under similar
conditions severely affected growth and develop-
ment of the transgenic plants (Shen et al. 2010).
A maize breeding programme for oil content
in kernels should be designed to circumvent
the decrease in grain yield. Understanding the
nature of quantitative genetic variation for oil
content in maize is essential to develop a suitable
breeding programme. Development of divergent
high-oil parental lines and their use in hybrid
development is the most important option for
development of high-oil maize. Favourable
82
alleles for high oil content can be assembled
through practising selection generation after gen-
eration accompanied by intermating of the
selected individuals or families. Though, this
will help in improving the genetic ceiling contin-
uously, but is a slow and time-taking process.
Efforts to accumulate the favourable QTLs in
the parental lines through marker-assisted recur-
rent selection (MARS) or development of nested
populations are needed. Transgenes like PINA,
PINB, ZmWRI1, and ZmLEC1 would be helpful
in further improving the oil content in the maize
inbreds and hence in hybrids. These different
approaches, if used in combination, may help in
speedy improvement of oil content in maize
populations or inbreds.
References
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Baud S, Lepiniec L (2010) Physiological and develop-
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associated with starch, protein, and oil deposition in
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for oil and protein percentage in maize. Proceedings of
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selection passes the century mark: a unique resource
for 21st century genomics. Trends Plant Sci
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DJ, Xiong H, Mai J, Screen SE, Val D, Lardizabal K,
Gruys K, Deikman J (2011) Expression of fungal
diacylglycerol acyltransferase2 genes to increase ker-
nel oil in maize. Plant Physiol 155:11461157
Pouvreau B, Baud S, Vernoud V, Morin V, Py C, Gendrot
G, Jean-Philippe P, Rouster J, Paul W, Rogowsky PM
(2011) Duplicate maize wrinkled1 transcription
factors activate target genes involved in seed oil bio-
synthesis. Plant Physiol 156:674686
Shen B, Allen WB, Zheng P, Li C, Glassman K, Ranch J,
Nubel D, Tarczynski MC (2010) Expression of
ZmLEC1 and ZmWRI1 increases seed oil production
in maize. Plant Physiol 153:980987
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QTL mapping of kernel oil concentration with high-
oil maize by SSR markers. Maydica 49:4148
Val LD, Schwartz SH, Kerns MR, Deikman J (2009)
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St. Paul
White PJ, Pollak LM, Duvick S (2007) Improving the
fatty acid composition of corn oil by using germplasm
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ML, Altosaar I, Giroux MJ (2010) The ectopic expres-
sion of the wheat Puroindoline genes increase germ
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Li J, Glassman K, Ranch J, Nubel D, Solawetz W,
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DGAT is a key determinant of oil content and compo-
sition in maize. Nat Genet 40:367372
 Maize Carotenoid Composition
and Biofortification for Provitamin A 7
Activity

Sandeep Kumar, Seema Sangwan, Rakesh Yadav,

Sapna Langyan, and Mohar Singh

Abstract
Carotenoids are fat-soluble antioxidant vitamin compounds derived
from the isoprenoid biosynthetic pathway. These natural pigments are
secondary metabolites and can be divided into two classescarotenes
and xanthophyllswhich play diverse biological roles in plants and
animals. Carotenoids with unsubstituted -ring end groups become
more important because of their provitamin A activity. Maize is the
third most staple food worldwide and also contains appreciable amount
of provitamin A carotenoids with wide range of genetic variability. This
makes it a good candidate crop for biofortification of provitamin A
carotenoids. The quantity of provitamin A carotenoids needed to allevi-
ate vitamin A deficiency (VAD) through biofortification depends upon
its bioavailability, which is influenced by a number of factors in an
individual. The bioavailability of biofortified maize can be known
through determining vitamin A equivalence. Recent advances have
shown that -carotene in biofortified maize has good bioavailability as
a plant source of vitamin A. So, a quantity of 15 g provitamin A g
1 dry
weight of kernel was targeted for biofortification. This chapter also
includes the carotenoid biosynthetic pathway, biofortification strategies,
recent advancements made toward biofortification of provitamin A, and
future perspectives.

S. Kumar (*)  M. Singh

Germplasm Evaluation Division, National Bureau
of Plant Genetic Resources, New Delhi, India
e-mail: kumarsandeep_biochem@rediffmail.com
S. Sangwan
Division of Microbiology, CCS HAU, Hisar, India
R. Yadav
Department of Bio and Nano Technology, GJUS&T,
Hisar, India
S. Langyan
Directorate of Maize Research, New Delhi, India
7.1 Introduction
Carotenoids are natural pigments derived from
the isoprenoid biosynthetic pathway and pro-
duced by most photosynthetic organisms that
have been shown to be beneficial to both plants
and animals. Carotenoids represent a diverse
group of more than 750 structures found in

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 83
DOI 10.1007/978-81-322-1623-0_7, # Springer India 2014
84
bacteria, fungi, algae, and plants (Britton et al.
2004). Of the carotenoids found in nature, 2050
are common in the human diet and about 20
are found in human blood and tissues (Johnson
2004). Biochemically, these secondary
metabolites are terpenoids and constitute a class
of fat-soluble antioxidant vitamin compounds
containing polyisoprenic structure. There are
two distinct classes of carotenoidscarotenes,
which contain only carbon and hydrogen, and
xanthophylls, which contain oxygen groups
(Van den Berg et al. 2000). Carotenoids are
lipophilic and are found in hydrophobic areas of
cells in close proximity to proteins and lipids
(Britton 1995; Van den Berg et al. 2000).
Carotenoids play various biological roles in
plants and animals. Some recent studies
suggesting that greater intakes of carotenoid-
containing foods result in reduced risks for sev-
eral chronic diseases have stimulated greater
interest in carotenoids (Canene-Adams et al.
2005). The exact chemical structure of individual
carotenoids is decisive for their biological
properties because it determines how they inter-
act with other molecules and integrates into
membranes (Rouseff et al. 1996).
Carotenoids can be divided into two groups,
provitamin A and non-provitamin A carotenoids,
depending upon their ability to release vitamin A.
Vitamin A deficiency has emerged as a serious
global health concern. A sustainable solution to
eliminate VAD is increasing the provitamin A
carotenoids in the major staple food crops, i.e.,
through biofortification. As maize is a staple
food worldwide and also rich in natural genetic
diversity for provitamin A carotenoids, qualify
as a suitable candidate crop for biofortification
(Wurtzel 2004). Recent advancements in areas of
genetics, biotechnology, biochemistry, and ana-
lytical tools have led to the identification of
QTLs, genes/alleles, or allozymes controlling the
regulatory steps of carotenoid biosynthetic and
utilizing pathways. Utilizing the available natural
genetic variability for provitamin A carotenoids,
maize breeders have succeeded in developing
biofortified maize lines containing 15 g -car-
otene/g dry kernel weight (Yan et al. 2010). So,
this chapter elaborates the importance of
S. Kumar et al.
carotenoids, various findings related to metabolic
pathway, developments, and future strategies
toward biofortification of maize.
7.2 Biosynthetic Pathway
The biosynthesis of carotenoids in plants occurs
on membranes of plastids with the regulating
enzymes encoded in the nuclear genes and
targeted to these plastids (Gallagher et al.
2004). Carotenoids are derived from the isopren-
oid biosynthetic pathway and are precursors of
the plant hormone abscisic acid and of other
apocarotenoids. The first committed step of this
pathway involves the condensation of two
geranylgeranyl pyrophosphate (GGPP) to form
15-cis-phytoene, a colorless C40 compound.
This constitutes the key regulatory step of the
pathway and is catalyzed by phytoene synthase
(PSY). Phytoene is converted to all-trans lyco-
pene (a red pigment) through a series of reactions
mediated by phytoene desaturase (PDS), -caro-
tene isomerase (Z-ISO), -carotene desaturase
(ZDS), and carotene isomerase (CRTISO).
The first branch point of this pathway occurs at
cyclization of lycopene cyclized by lycopene -
cyclase (LCYE) and/or lycopene -cyclase
(LCYB) to generate - and -carotenes, respec-
tively. Relative activities of LCYB and LCYE
are hypothesized to regulate the proportion of
carotenes directed to each branch of this path-
way. - and -carotenes are subsequently
hydroxylated and modified to form the various
xanthophylls. The carotenoid biosynthetic path-
way in plants is shown in Fig. 7.1.
7.3 Biological Roles
In plants and animals, these pigments play
diverse biological roles as mentioned below:
As natural pigments, carotenoids are responsi-
ble for different colors present in plant parts,
which is determined by the number and location
of the double bonds present within the structure
(Watson 1962). As accessory pigments in the
photosynthetic apparatus, they play various roles
7 Maize Carotenoid Composition and Biofortification for Provitamin A Activity 85

Fig. 7.1 Carotenoid Geranylgeranyl pyrophosphate

biosynthetic pathway (GGPP)
in plants
Phytoene synthase
Phytoene
Phytoene desaturase
-carotene isomerase
-carotene
-carotene desaturase
Carotene isomerase

All-trans lycopene
Lycopene -cyclase Lycopene -cyclase
Lycopene -cyclase Lycopene -cyclase
-carotene -carotene
-carotene hydroxylase -carotene hydroxylase

Zeinoxanthin -cryptoxanthin

-carotene hydroxylase -carotene hydroxylase

Lutein Zeaxanthin
Violaxanthin de-epoxidase Zeaxanthin epoxidase
Antheraxanthin
Violaxanthin de-epoxidase Zeaxanthin epoxidase
Violaxanthin

Abscisic acid

such as in light harvesting and photoprotection, as
attractants for seed dispersal and pollination, and
as precursors of some scents as antioxidants
(Cunningham 2002; Fraser and Bramley 2004;
Howitt and Pogson 2006). In spite of various
biological functions, their role as antioxidants
appears to be ubiquitous, which is best understood
in chloroplasts, where desaturated carotenoids
quench triplet chlorophyll and singlet oxygen,
preventing the formation of reactive oxygen spe-
cies and photo-oxidation of the contents of the
organelle (Niyogi 1999). Carotenoids also act as
precursors to various cleavage products, such as
the apocarotenoid abscisic acid (ABA), which
regulates plant growth, embryo development, dor-
mancy, and stress responses (Nambara and
Marion-Poll 2005), and additional apocarotenoid
products, such as strigolactone and others, whose
functions are still not known but are considered
essential elements that may affect plant yield
(Booker et al. 2004; Akiyama and Hayashi
2006). Carotenoid antioxidants protect
membranes from lipid peroxidation under heat
and light stress conditions (Havaux et al. 2007;
Johnson et al. 2007).
Provitamin A activity is the classical
biological function of carotenoids in mammalian
systems (USDA 2008). Carotenoids such as -
carotene, -carotene, and -cryptoxanthin have
provitamin A activity as these contains
unsubstituted -ring end groups. Among these,
-carotene contains two provitamin A structures,
i.e., two non-hydroxylated -ionone rings, in
comparison to -cryptoxanthin and -carotene,
86
which contains single non-hydroxylated -
ionone ring, so it has twice the activity of the
others. In case of mammals, provitamin A
carotenoids are cleaved in the intestinal lumen
to produce retinal (vitamin A). In addition,
xanthophylls such as lutein and zeaxanthin are
the essential components of the macular pigment
of the eye (Beatty et al. 1999). Macular pigment
protects retinal rods and cones by filtering harm-
ful UV/blue light wavelengths (Mares-Perlman
and Klein 1999; Kopsell and Kopsell 2006).
Apart from these activities, several other essen-
tial biological functions of carotenoids, particu-
larly in human health, include reducing the risk
of degenerative diseases such as cardiovascular,
cancers, cataract, and muscular degeneration via
their role as antioxidants and/or as regulators of
the immune system (Fraser and Bramley 2004;
Johnson 2004; Tang et al. 2005).
Economic benefits to animal production
systems are also associated with carotenoids.
Diets of chickens with high levels of carotenoid
pigments have also been associated with a desir-
able color of the egg yolks and of the skin of
broilers and fryers (Blessin et al. 1963; Perez-
Vendrell et al. 2001). -carotene levels may work
synergistically with other fat-soluble vitamins in
the protection of broiler leg meat from oxidation
(Ruiz et al. 1999). This coloration of yolks and
skin is perceived as associated with good health
and quality by some consumers (Hadden et al.
1999). The industrial use of carotenoids involves
their application for nutrient supplementation,
pharmaceutical purposes and as food colorants
and animal feeds (Weber 1987).
7.4 Implications of Vitamin A
Deficiency
Provitamin A activity is the classical biological
function of carotenoids in mammalian systems
as only its deficiency in the body may result
into causal factor for numerous other diseases.
It is well known that adequate vitamin A
intake is important for vision, growth, cellular
S. Kumar et al.
differentiation and proliferation, reproduction,
and the integrity of the immune system
(Sommer 1982). Globally, approximately one-
third of preschool-age children and 15 % of
pregnant women are estimated to be vitamin A
deficient (WHO 2009). The problem becomes
more severe particularly in the developing
countries whose poor populations rely on a
single staple crop for their sustenance, e.g.,
Africa and Southeast Asia have the highest
burden of vitamin A deficiency (WHO 2009).
The consequences of VAD include blindness,
reduced growth in children, and increased mor-
bidity and mortality (Sommer and West 1996;
Shankar et al. 1999; Rice et al. 2004; Maida
et al. 2008).
7.5 Maize Carotenoid Composition
Maize exhibits considerable natural variation
for kernel carotenoids, with some genotypes
accumulating as high as 66.0 g/g (Harjes
et al. 2008). Yellow maize kernel carotenoids
are present in different isoforms, including two
carotenes, - and -carotene, and three
xanthophylls, -cryptoxanthin, zeaxanthin, and
lutein (Watson 1962; Weber 1987). The pre-
dominant carotenoids in maize kernels, in
decreasing order of concentration, are lutein,
zeaxanthin, -carotene, -cryptoxanthin, and -
carotene. Generally, provitamin A carotenoids
constitute only 1020 % of total carotenoids in
maize, whereas zeaxanthin and lutein each com-
monly represent 3050 %. The amounts of pro-
vitamin A in traditional yellow maize varieties
range from 0.25 g to 2.5 g g
1 dry weight.
In typical maize, concentrations of provitamin
A carotenoids, i.e., -carotene, -carotene, and
-cryptoxanthin, range from 0 to 1.3, 0.13 to 2.7,
and 0.13 to 1.9 nmol/g, respectively (Kurilich and
Juvik 1999). Although -carotene has the highest
provitamin A activity, it is present in a relatively
low concentration (0.51.5 g/g) in most yellow
maize grown and consumed throughout the world
(Harjes et al. 2008).
7 Maize Carotenoid Composition and Biofortification for Provitamin A Activity
7.6 Biofortification
Biofortification is the development of
micronutrient-dense staple crops using the best
traditional breeding practices and modern bio-
technology. In developing countries, staple food
crops predominate in the diets of the poor, so,
biofortification inherently targets the most vul-
nerable populations (Bouis 2003). Bioforti-
fication differs from ordinary fortification
because it focuses on making plant foods more
nutritious as the plants are growing, rather than
having nutrients added to the foods when they
are being processed. The biofortified seeds can
be easily reproduced by poor farmers, and thus
the seeds are a sustainable means to target remote
rural communities not served by conventional
seed markets (Qaim et al. 2007).
To alleviate the vitamin A deficiency from
rural areas of developing countries, bioforti-
fication of major staple food crops of respective
areas or countries with provitamin A carotenoids
is the only feasible way, since this will better
ensure the targeting and compliance. Further-
more possible economic benefits to animal pro-
duction systems are also associated with
carotenoids, which can be increased through
biofortification for carotenoids.
As the third most important cereal staple food
crop worldwide (FAPRI 2009), a major cereal
staple for African consumers (FAOSTAT
2010), maize has also the advantage of being
the only crop that contains appreciable amount
of carotenoids (Wurtzel 2004) with wide range of
genetic variability. The first solid food given to
African infants includes white maize porridges
devoid of provitamin A (Faber et al. 2005;
Poggensee et al. 2004) and are also widely con-
sumed by older children and adults. The
nutritional improvement of maize with provita-
min A carotenoids would have a significant
impact on the target populations. Utilizing
the available natural genetic variability for pro-
vitamin A carotenoids, maize breeders have
succeeded in developing biofortified maize lines
containing 15 g -carotene/g dry kernel
weight (Yan et al. 2010).
87
Before devising a strategy to enhance the pro-
vitamin A carotenoids in maize, one question
that must be addressed is how much quantity is
needed to alleviate VAD. This is related to the
bioavailability or the fraction of an ingested
nutrient that becomes available to the body for
utilization in physiological functions or for stor-
age (Jackson 1997; Fraser and Bramley 2004).
There are numerous factors that influence bio-
availability, including nutrient status of the host,
species of carotenoid, food matrix, and amount
of food consumed in the meal. In biofortified
maize, the bioefficacy of the -carotene can be
predicted by determining experimentally its vita-
min A equivalence. The -carotene quantity
needed to provide vitamin A activity equivalent
to 1 g of retinol is known as vitamin A equiva-
lence. The extent of conversion of -carotene to
vitamin A is highly variable among well-
nourished people. A recent study evaluated the
effect of genetic polymorphisms in the BCMO1
gene that encodes -carotene 15,150 -
monooxygenase on the conversion of -carotene
to retinyl palmitate (Leung et al. 2009). Taking
these factors into account, nutritionists have
estimated that 15 g provitamin A g
1 dry weight
of kernel could greatly alleviate vitamin A defi-
ciency (www.harvestplus.org). Li et al. (2010)
carried out an experiment on women to quantify
the vitamin A equivalence of the -carotene in -
carotene-biofortified maize based on consumption
of a single serving of maize porridge and found
that -carotene in biofortified maize has good
bioavailability as a plant source of vitamin A.
So, improving the micronutrient balance of
staple crops such as maize through bioforti-
fication is therefore an economically and socially
sound way to address micronutrient malnutrition,
including VAD, on a global scale (Tanumihardjo
et al. 2008).
7.7 Strategies for Biofortification
of Maize
To increase the provitamin A potential of maize,
there is a need to maximize biosynthetic
flux toward carotenoid synthesis, limit the
88
carotenoids degradation, inhibit branching points
leading to non-provitamin A carotenoids and
inhibit enzymes leading to the conversion of
provitamin A to non-provitamin A carotenoids.
This can be achieved by regulating the rate-
limiting steps of the pathway or identifying the
alleles contributing toward synthesis of provita-
min A carotenoids as mentioned below.
The enzyme phytoene synthase catalyzes the
first rate-limiting step of the carotenoid biosyn-
thetic pathway and is the most important as its
activity controls the carbon flux toward caroten-
oid biosynthesis. The first branch point of this
pathway occurs at cyclization of lycopene where
action of lycopene beta cyclase (LCYB) at both
ends of linear lycopene produces a molecule with
two -rings. Alternatively, the coaction of LCYB
and lycopene epsilon cyclase (LCYE) generates
a , -carotene that is a precursor to lutein.
Relative activities of LCYB and LCYE are
hypothesized to regulate the proportion of
carotenes directed to each branch of this path-
way. If somehow flux can be shifted toward
more synthesis of -carotene, it will be a boost
for provitamin A synthesis. But as the concentra-
tion of provitamin A carotenoids increases, a
large amount is hydroxylated to -cryptoxanthin
(-CX) and zeaxanthin (Z), which have 50 % and
0 % of the provitamin A activity of -carotene,
respectively. So, there is need to downregulate
the activities of carotene hydroxylases. Further-
more the inhibition of the steps leading to degra-
dation of the carotenoids or directing toward
other metabolic pathways such as synthesis of
ABA will also help in increasing the concentra-
tion of provitamin A.
7.8 Recent Advancements Related
to Metabolic Pathways
Toward Increasing Provitamin
A Carotenoids
Use of composite interval mapping led to identi-
fication of two candidate genes, yellow 1 and
viviparous 9, which may be responsible for quan-
titative variation in carotenoids. The yellow 1
gene is associated with phytoene synthase, the
S. Kumar et al.
enzyme catalyzing the first dedicated step, and
the viviparous 9 gene is associated with zeta-
carotene desaturase, an enzyme catalyzing an
early step, in the carotenoid biosynthetic path-
way (Wong et al. 2004). Similarly, Chander et al.
(2008) constructed a genetic linkage map using a
233 recombinant inbred lines derived from a
cross between By804 and B73 and identified 31
putative QTL in total including 23 for individual
and 8 for total carotenoids. Two loci, i.e., y1 and
y9, that explained most of the phenotypic varia-
tion in carotenoids contents were identified along
with a gene-targeted marker (Y1ssr) in the can-
didate gene phytoene synthase 1 (psy1) tightly
linked to a major QTL explaining 6.627.2 %
phenotypic variation for levels of carotenoids.
They also emphasize the role of QTL cluster
located at y9 locus for pyramiding favorable
alleles controlling contents of carotenoids from
diverse maize germplasm. Later on, Chen et al.
(2010) found that maize y9 locus encodes -car-
otene isomerase (Z-ISO), an enzyme necessary
for endosperm carotenogenesis in plants.
Another catabolic gene affecting seed
carotenogenesis, i.e., CCD1 from maize, was
cloned and found to cleave carotenoids effec-
tively (Sun et al. 2008; Vogel et al. 2008). The
position of ZmCCD1, chromosome 9.07, was
found linked to the dominant white cap 1 (wc1)
locus involved in the depletion of endosperm
carotenoids through gene dosage effect
(Vallabhaneni et al. 2010).
Several haplotypes of the gene encoding lyco-
pene epsilon cyclase (lcyE) that substantially
increase the ratio of - to -carotenoids in
maize grain were identified by Harjes et al.
(2008). To find out the correlation between carot-
enoid content and candidate gene transcript
levels, a maize germplasm collection was used
by Vallabhaneni and Wurtzel (2009), and it was
observed that transcript levels of paralogs
encoding isoprenoid isopentenyl diphosphate
and geranylgeranyl diphosphate-producing
enzymes, DXS3, DXR, HDR, and GGPPS1,
were positively correlated with endosperm carot-
enoid content. PSY1 and CrtISO transcripts were
found to be positively and inversely correlated,
respectively, for carotenoid pathway enzymes.
7 Maize Carotenoid Composition and Biofortification for Provitamin A Activity
Furthermore, the ZEP (zeaxanthin epoxidase)
transcript level, the enzyme involved in the
depletion of carotenoid pool for its conversion
to ABA, was also examined, and carotenoid
accumulation was found inversely associated
with ZEP1 and ZEP2 transcript levels.
Using metabolite sorting on maize diversity core
collection, Vallabhaneni et al. (2009) identified the
enzyme carotene hydroxylase encoded by the
Hydroxylase3 (HYD3) locus, whose transcript
levels were negatively correlated with high -caro-
tene levels and positively correlated with zeaxan-
thin levels. Using PCR genotyping of 51 maize
lines, they also showed that the HYD3 locus could
explain 36 % variation and fourfold difference in -
carotene levels. Yan et al. (2010) demonstrated
through association and linkage population studies
in maize that the gene encoding -carotene hydrox-
ylase 1 (crtRB1/HYD3) underlies a principal quan-
titative trait locus associated with -carotene
concentration and conversion in maize kernels.
crtRB1 alleles associated with reduced transcript
expression were found correlated with higher -
carotene concentrations.
7.9 Progress Made Toward Maize
Biofortification for Provitamin
A Carotenoids
In the past few years, significant progress has
been made toward maize biofortification. Con-
ventional breeding has led to the development of
a few high -carotene maize lines having a max-
imum of 13.6 g g
1 of provitamin A g
1 dry
weight kernel that approach the target of
15 g g
1 of provitamin A g
1 dry weight kernel
(Kurilich and Juvik 1999; Islam 2004; Harjes
et al. 2008). Biofortified maize lines containing
15 g -carotene/g dry kernel weight have been
successfully developed by the breeders (Yan
et al. 2010). Now these identified source lines
are being routinely used as parents for new
crosses to obtain new sources with greater provi-
tamin A carotenoids. As breeding programme is
cyclic in nature, there is need of continuous
screening of germplasm and to exploit the natural
variability available through molecular marker-
89
assisted selection (MAS) for biofortification
purpose.
Transgenic strategies provide important tools
to transfer the desired trait from one species to
another. Aluru et al. (2008) show that maize
seeds can be metabolically engineered to pro-
duce high levels of provitamin A comparable to
50 % EAR values by overexpressing the bacterial
crtB and crtI genes in an endosperm-specific
manner, using a modified and highly active c-
zein promoter. As maize exhibit considerable
natural variation for provitamin A carotenoids,
transgenic approaches have not been used for its
biofortification.
7.10 Future Perspectives
Maize biofortification for provitamin A
carotenoids benefits human health and also adds
to commercial value of food as these are natural
colorants. National germplasm collections hold
untapped potential for maize improvement.
Biofortification of maize requires large-scale
germplasm screening and utilization of identified
high provitamin A carotenoid material into the
breeding programme. There is need to identify
more QTLs through population development,
whereas mutational studies can provide an
insight about the regulatory points. Similarly,
stage-specific metabolite profiling and its corre-
lation to candidate gene expression will provide
important information regarding regulation
chemistry and expression patterns. So, an inter-
disciplinary approach including biochemistry,
genetics, plant breeding, and nutrition is required
for understanding and identifying more QTLs,
the rate-limiting steps of the pathway, gene
expression patterns with respect to time,
allozymic diversity, etc., which, in turn, might
provide important information for deciding
futuristic strategies. Furthermore, the developed
biofortified maize material should not only con-
tain higher quantities of provitamin A
carotenoids but also have all other crucial traits
including higher yield, insect-pest and disease
resistance, and better nutritional quality.
90
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 Part IV
Biotic and Abiotic Stresses in Maize
 Insect-Pests and Their Management:
Current Status and Future Need 8
of Research in Quality Maize

M.K. Dhillon, V.K. Kalia, and G.T. Gujar

Abstract
Maize (Zea mays L.) is an important staple food for millions of people
across the world. However, different stress factors mainly the insect-
pests, viz., maize stalk borer, pink stem borer, sugarcane leafhopper,
shoot bug, armyworm, shoot fly, corn leaf aphid, cob borer, and termites,
have constrained the increase in yield potential of the maize genotypes
deployed in India. The hybrid initiative turned out to be exceedingly
important for Quality Protein Maize (QPM) efforts, and many countries
in the developing world including India are becoming increasingly
interested in QPM to reduce malnourishment and to sustain nutritional
security. However, it is also likely that the increase in concentration and
quality of nutritional components particularly the protein in QPM might
favor the proliferation of insect-pests as they too prefer quality food for
their growth and development and could be major constraints to increas-
ing production and productivity of QPM. Therefore, development and
deployment of high-yielding and insect-resistant QPM genotypes under
the umbrella of integrated pest management system might help in
narrowing down the yield gap by reducing the crop losses caused by
insect-pests. Since maize is damaged by an array of insect groups with
different feeding habits right from the seedling stage to maturity of the
crop, no single strategy is sufficient to manage such complex group of
insect-pests. Damage potential of different insect-pests, status of host
plant resistance and the mechanisms of resistance involved, and the
management of major insect-pests have also been discussed in this
chapter.

8.1 Introduction

Maize (Zea mays L.) is the third most important

M.K. Dhillon (*)  V.K. Kalia  G.T. Gujar
Division of Entomology, Indian Agricultural Research
Institute, New Delhi 110012, India
e-mail: mukeshdhillon@iari.res.in;
mukeshdhillon@rediffmail.com
food crop after rice and wheat and is an important
staple food for millions of people in Asia,
Africa, and Latin America. According to recent
estimates, the maize is being cultivated over an

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 95
DOI 10.1007/978-81-322-1623-0_8, # Springer India 2014
96
area of 8.12 million hectare (mha) with annual
production of 19.77 million tons (mt),
contributing 49 % to poultry feed, 25 % to
human food, 12 % to animal feed, 12 % to indus-
trial products mainly the starch, and 1 % each in
brewery and seed (Dass et al. 2008). Globally,
about 200 million children younger than 5 years
are undernourished for protein, leading to a num-
ber of health problems including stunted growth,
weakened resistance to infection, and impaired
intellectual development. Several million people,
particularly in the developing countries, derive
their protein and calorie requirements from
maize. The maize grain accounts for about
15 %56 % of the total daily calories in diets
of people in about 25 developing countries, par-
ticularly in Africa and Latin America. Although
maize has contributed significantly (8 %) to the
food basket in India, the nutritional quality of
maize protein in the traditional genotypes is
poor because of imbalanced amino acid compo-
sition due to deficiencies of two main essential
amino acids, lysine and tryptophan, and excess of
leucine. The discovery of association of high
lysine and tryptophan with opaque-2 (o-2)
maize endosperm by Mertz and his group in
1964 opened up new vistas in improving the
protein quality of maize. Through the combined
use of o-2 gene and genetic modifiers, the QPM
gene pools have been improved for yield and
associated traits, kernel modification, and
appearance and the QPM versions have
performed quite similar to normal counterparts
in yield and agronomic traits (Vasal et al. 1984,
1994; Vasal 2002). The discovery of another
mutation, floury-2 (fl2), also has the ability to
alter endosperm nutritional quality (Bjarnason
and Vasal 1992). However, o-2 and fl2 genes
make the QPM endosperm soft and attractive to
the stored grain pests. Furthermore, the hybrid
initiative turned out to be exceedingly important
for QPM efforts and success, and many countries
in the developing world, including India, are
becoming increasingly interested in it. It is also
not surprising to experience increased insect-
pest problems under field as well as storage
conditions with the deployment of QPM
genotypes, as the insects too prefer quality food
M.K. Dhillon et al.
for their growth and development and could be
major constraints to increasing production and
productivity of QPM.
The grain yields of traditional maize genotypes
in India are quite low, and in spite of sincere
efforts, the yield potential of maize genotypes
deployed in India has not reached even half to
that in the United States. Furthermore, the gap in
yield potential of maize genotypes under experi-
mental conditions and that under farmers fields
is huge, due to different stress factors, mainly
the insect-pests during vegetative and storage
conditions, and needs greater attention to break
this yield gap. The maize is attacked by about
139 insect-pests with varying degree of damage
under field and storage conditions. Among the
field pests, maize stalk borer [Chilo partellus
(Swinhoe)], pink stem borer [Sesamia inferens
(Walker)], sugarcane leafhopper [Pyrilla perpu-
silla (Walker)], shoot bug [Peregrinus maidis
(Ashmead)], armyworm [Mythimna separata
(Walker)], shoot fly [Atherigona spp.], corn leaf
aphid [Rhopalosiphum maidis (Fitch)], cob borer
[Helicoverpa armigera (Hubner)], and termites
[Macrotermes spp. and Odontotermes spp.] are
some of the major yield-reducing factors of
maize right from seedling emergence to harvest.
Maize weevils, Sitophilus oryzae (L.) and
Sitophilus zeamais Motschulsky, cause serious
grain losses both under field and storage conditions
in the tropical countries of the world. The maize
stalk borer, C. partellus alone, causes 26.780.4%
yield loss under different agroclimatic conditions
in India (Reddy and Zehr 2004). Host plant resis-
tance is one of the effective means of minimizing
losses due to insect-pests. However, most of the
maize varieties and hybrids released for cultivation
are susceptible to C. partellus during vegetative
stage (Kumar 1997) and maize weevils, Sitophilus
spp., under field and storage conditions (Arnason
et al. 1993; Hossain et al. 2007). Screening of
maize germplasm for resistance to spotted stem
borer, C. partellus, has led to identification of
several maize lines with tolerance to this pest
(Kanta et al. 1997), and several morphological,
anatomical, and biochemical factors have
been reported to be associated with resistance/sus-
ceptibility to stem borer in maize (Kumar 1997;
8 InsectPests and Their Management: Current Status and Future. . .
Bhanot et al. 2004). However, the success in
incorporation of insect resistance in agronomically
elite cultivars has been very slow, as it has been
difficult to monitor all the traits in breeding
populations, and incorporation of yield and quality
traits along with insect resistance has not been
rewarding in terms of yield advantage. Deploy-
ment of high QPM genotypes with resistance to
insect-pests and superior yield could be one of the
most effective, economic, socially acceptable, and
environmentally safe tool for increasing maize
productivity. Major insect-pests and their manage-
ment in maize have also been discussed in this
chapter.
8.2 Status of Host Plant Resistance
to Insect-Pests in Maize
Considerable genotypic variation in grain protein
content has been found in both QPM and non-
QPM cultivars; however, protein concentration
in the grain decreases with the increase in grain
yield (Ortiz-Monasterio et al. 2001; Worku et al.
2007). The zein synthesis can be manipulated by
nitrogen fertilization and genetic means, and a
positive relationship between the zein content
and grain yield needs to be established (Vasal
2001). Thus, it is possible to develop N-efficient
cultivars that may combine high yield potential
and protein quality at all levels of soil fer-
tility (Worku et al. 2007). Undesirable
characteristics such as reduced yield than normal
maize, low grain consistence, and a farinaceous
endosperm that retains water resulting in a soft,
chalky endosperm that upon drying makes it
more prone to insect damage and also affects
harvest ability (Singh and Venkatesh 2006).
The maize weevil, S. zeamais, is an important
pest of maize in the tropical countries of the
world (Arnason et al. 1993), while maize weevil,
S. oryzae, is major pest of maize under field and
storage conditions in India. Several weevil-
tolerant QPM lines have been identified, which
can be used in breeding for resistance to this pest
(Hossain et al. 2007). Stem borers are regarded as
a major limiting factor to the production of maize
in India (Panwar 2005), of which the spotted
stem borer, C. partellus, is the most serious and
97
prevalent pest of maize in different agroclimatic
regions of the country. Screening of maize germ-
plasm for resistance to spotted stem borer, C.
partellus, has reported low to moderate levels
of resistance to this pest (Kanta et al. 1997;
DMR 2007; Rakshit et al. 2008; Sekhar et al.
2008). Several new sources of insect resistance
have also been identified and supplemented to
the existing resistance sources against corn
earworm, Helicoverpa zea (Boddie); corn borer,
Ostrinia nubilalis (Hubner); sugarcane borer,
Diatraea grandiosella (Dyar); and fall army-
worm, Spodoptera frugiperda (J.E. Smith) in
maize, but most of the maize varieties and
hybrids released for cultivation are susceptible
to spotted stem borer, C. partellus (Kumar
1997). The mechanisms, inheritance, nature of
gene action, and application of molecular
markers for resistance to several insect-pests
such as maize borers, viz., D. grandiosella, D.
saccharalis (Fabricius), Sesamia nonagrioides
(Lefebvre), and O. nubilalis (Williams et al.
1995, 1998; Kumar and Mihm 1996; Khairallah
et al. 1997; Cartea et al. 2001; Butron et al. 2005;
Krakowsky et al. 2007); corn earworm, H. zea
(Widstrom and Snook 2001), and corn leaf aphid,
R. maidis (Fitch) (Bing and Guthrie 1991), are
well understood; however, such information is
poorly studied for C. partellus in maize. It has
been difficult to breed for resistance to stem
borers, despite the heavy losses incurred year
after year. Moreover, screening for resistance to
stem borers in maize has not been used consis-
tently to gauge levels of resistance under con-
trolled greenhouse and field conditions, which is
prerequisite for host plant resistance studies.
Screening for resistance to stem borers under
greenhouse and field conditions using artificial
infestation has reported to be the most effective
method of characterizing insect-resistant
cultivars (Sharma et al. 2007).
8.3 Mechanisms of Resistance
in Maize to Insect-Pests
The total biochemical energy in crop plants is
fixed, and the changes in biochemical
constituents through manipulation of metabolic
98
pathways happen to be at the cost of change in
the concentration of one another. Biochemical
mechanism of insect defense in crop plants is
mainly governed by constitutive and/or induced
plant metabolic compounds. This is in the con-
text that increases in concentration and quality of
nutritional compounds particularly the protein in
QPM might favor the proliferation of insect-pests,
as the insects too prefer quality food for their
growth and development and could be major
constraints in increasing production and produc-
tivity of QPM. Several morphological and
anatomical plant characters such as seedling
vigor, leaf width, trichome density, plant height,
length of internode, tassel ratio, stem thickness
and hardness, and lignified vascular bundles have
been reported to be responsible for resistance/sus-
ceptibility to insect-pests in maize (Durbey and
Sarup 1982; Kumar and Saxena 1985; Kumar
1997; Rao and Panwar 2000, 2001; Ashfaq and
Farooq-Ahmad 2002; Bhanot et al. 2004). Hairy
leaf surface was found to be negatively correlated
with oviposition by C. partellus (Kumar 1992; van
den Berg 2006). In addition, numerous biochemi-
cal factors such as nitrogen, phosphorus, potash,
silica, iron, sugars, amino acids, protein content,
tannins, and polyphenols have also been reported
to be associated with resistance/susceptibility to
insect-pests in maize (Kumar and Saxena 1985;
Kabre and Ghorpade 1997, 1999; Kumar 1997;
Bhanot et al. 2004). Biochemical constituents,
viz., dotriacontanol, heptadecanol, and nona-
decanol, have been reported to be associated with
oviposition deterrence by C. partellus females on
maize (Varshney et al. 2003) and certain chemicals
of resistant genotype extractable in hexane,
elicited larval repulsion (Varshney et al. 2007).
The presence of mixtures of aliphatic aldehydes,
ketones, alcohols, esters, as well as terpenoids and
other aromatic compounds in the volatile
emissions of corn leaves has been reported to be
associated with orientation and ultimate recogni-
tion of host plants by the insects (Buttery and Ling
1984), and six plant volatiles, viz., octanal,
nonanal, linalool, naphthalene, allylanisole, and
eugenol, have been reported to mediate host loca-
tion and oviposition by C. partellus females
(Khan et al. 2000; Birkett et al. 2006).
M.K. Dhillon et al.
Very young corn seedlings govern high level of
resistance to the European corn borer (ECB)
because of synthesis of high concentrations of
6-methoxybenzoxazolinone (6-MBOA) and
2,4-dihydroxy-7-methoxy-1,4-benzoxazine-3-one
(DIMBOA) during this stage (Beck 1965). Later
on, Klun and Brindley (1966) found that 6-MBOA
is not present in vivo but its precursora gluco-
sideis present when the corn seedling tissue
is crushed by insect mandibles, which rapidly
hydrolyzed to glucose and aglucon DIMBOA,
and the DIMBOA is then converted into 6-
MBOA by enzymatic action. Furthermore, Klun
et al. (1970) reported that the high concentration
of DIMBOA in young corn plants imparts resis-
tance against ECB, and as plant grows older, the
DIMBOA starts decreasing in the stalk whorl
leaves and get concentrated in roots. DIMBOA
has also been found responsible for resistance
against corn earworm, H. zea. These secondary
metabolites are produced independent of the pres-
ence of pest in a tissue. In addition to secondary
metabolites, ubiquitous phenolic acids, especially
ferulic acid, may contribute to insect resistance in
maize. The biosynthesis of these defense-related
metabolites has a common root in the shikimic
acid pathway. Maysin, chlorogenic acid, and phe-
nolic acids originate from the phenylalanine
branch of the pathway, and biosynthesis of
benzoxazinoids shares intermediates with the
tryptophan metabolism (McMullen et al. 2009).
Later, it was also found that flavone glycoside and
maysin in the maize silk govern antibiosis mecha-
nism of resistance to H. zea larvae.
A blend of volatile compounds is being emitted
by the herbivore-damaged maize plants, and the
natural enemies also use them as chemical cues
for spotting the host insects on the damaged
plants. In response to foliar damage by lepidop-
teran larvae in maize, the plant releases a complex
mixture of volatiles, which attract females of the
parasitic wasp, Cotesia marginiventris (Cresson),
to the damaged maize plant to oviposit on
the insect larvae (Turlings et al. 1990, 1991).
The parasitized larvae consume less plant material
and will die upon emergence of the parasitoid,
which can benefit the plant (Hoballah and
Turlings 1999; Hoballah et al. 2004). The
8 InsectPests and Their Management: Current Status and Future. . .
composition of the lepidopteran-induced volatiles
varies between different lines of maize and teo-
sinte (Gouinguene et al. 2001; Degen et al. 2004)
and is influenced strongly by abiotic factors like
temperature, light intensity, and nutritional status
of the plant (Gouinguene and Turlings 2002). The
phenomenon of tritrophic interactions involving
maize, herbivores, and enemies was first
demonstrated by Turlings et al. (1990) and after-
wards reported in more than 15 plant species
(Dicke 1999; Kessler and Baldwin 2002). The
attraction of herbivore enemies has been shown
to benefit the plants by reducing subsequent her-
bivory and increasing reproductive fitness
(Hoballah and Turlings 1999; Van Loon et al.
2000; Kessler and Baldwin 2001), although such
advantages have not been realized in all the cases.
The maize volatile blend consists of indole,
products of the lipoxygenase pathway, and a
large number of mono- and sesquiterpenes
(Kollner et al. 2004). Attempts to identify the
compounds that are crucial for the attraction
of parasitic wasps have been hampered by the
complexity of blends and the difficulty of
obtaining individual compounds with the correct
chirality for bioassay (DAlessandro and Turlings
2005, 2006). Although several physicochemical
traits have been reported to be associated with
insect resistance, their use in selection of insect-
resistant cultivars has been underutilized.
8.4 Major Insect-Pests of Maize
In India, maize productivity is very low in com-
parison to its potential. The most important rea-
son for its low productivity is improper
management of the crop and insect-pests infes-
tation. The maize is damaged by an array of
insect-pest groups, viz., soil dwellers, foliage
feeders, stem borers, flower feeders, and cob
feeders under field conditions, and rice weevil,
Khapra beetle, lesser grain borer, and grain moth
under storage conditions (Kumar et al. 2001).
The insect-pests status too is under continuous
change in both inter- and intra-agroclimatic
conditions due to unsuitable agroclimatic
conditions, cultivation of high-yielding but
99
insect-susceptible hybrids or varieties, indiscrim-
inate use of pesticide, reduction in natural
enemies population, use of excess dose of
fertilizers, change in cropping pattern, availabil-
ity of alternate hosts, and increase in maize grow-
ing throughout year. The important insect-pests
of maize are described below:
Maize stalk borer (C. partellus): Among all
the insect-pests of maize, stalk borer is the
most serious pest found throughout India.
The yield losses due to this pest have been
recorded up to 3580 %. Sowing time
influences the stalk borer attack as crop sown
during the first fortnight of March shows
higher infestation than that sown in the second
fortnight of February or the second fortnight
of June and October. The stem borer infesta-
tion ranges between 4.7 % and 24.0 % in
summer (February to March) and 9.5 and
37.4 % in winter (November to December).
Higher maximum temperature (>13  C) and
relative humidity (>70 %) favor for stem
borer attack, and every 1  C reduction in min-
imum temperature decreases 1.7 % stem borer
damage at 30 days after sowing.
Pink stem borer (S. inferens): This is a major
pest of wheat and also causes serious losses in
maize. Infestation of this pest has increased
gradually in North India and causes consider-
able yield losses in maize, while the losses
due to this pest in South India are compara-
tively higher (2580 %).
Shoot fly (Atherigona spp.): Shoot fly is a seri-
ous pest of maize in spring maize in northern
India. It attacks the crop at the seedling
stage. Three species of shoot fly are found in
the country, of which most prominent are
Atherigona naqvi Steyskal (in Ludhiana,
Punjab), Atherigona orientalis Schiner (in
Pantnagar), and Atherigona soccata Rondani
and A. naqvi (in Delhi). Maximum temperature
of 3034  C, minimum temperature of
2425  C, and relative humidity of 4075 %
are favorable for severe attack, while for mini-
mum incidence, maximum temperature of
2425  C, minimum temperature of 2122  C,
and relative humidity of 8086 % in the second
and third week after sowing are favorable.
100
Cutworms/armyworms: Severe infestation of
cutworms can be seen in Rabi maize mainly in
the Terai area of Uttar Pradesh and Uttarakhand
where it causes loss throughout the year, which
has now extended to Bihar, Delhi, Haryana,
Rajasthan, Madhya Pradesh, West Bengal,
Orissa Punjab, and Jammu and Kashmir in all
seasons of the crop. The armyworm damage was
earlier confined mainly to northern India on
Rabi maize, which has also now been recorded
to damage Kharif maize of Himachal Pradesh,
Punjab, Karnataka, Tamil Nadu, Maharashtra,
Manipur, Orissa, Gujarat, and Uttar Pradesh.
Sugarcane leafhopper (P. perpusilla): This is a
serious pest of sugarcane, which has now been
recorded infesting Kharif maize in the north-
ern states of India mainly Punjab, Haryana,
Maharashtra, Uttar Pradesh, and Uttarakhand.
Indiscriminate use of insecticides and change
in climatic conditions causing heavy reduc-
tion in parasitoids of hopper might have
increased severity of the pest during recent
years.
Shoot bug (P. maidis): Adults and nymphs
suck sap, resulting in unhealthy, stunted, and
yellow plants. The leaves whither from
top downwards, and plants die if attack is
severe. Honeydew excreted by shoot bug
causes growth of sooty mold on leaves,
midribs of leaves turn red due to egg laying
and may dry subsequently. Shoot bug is
occasionally a serious pest on maize. It has
been found attacking maize in South India and
Madhya Pradesh with peak activity during
AugustOctober.
Corn leaf aphid (R. maidis): The aphid colonies
may completely cover emerging tassels and
the surrounding leaves, preventing pollen
release. This insect also acts as a vector of
the sugarcane mosaic virus, maize dwarf
mosaic virus, and maize leaf fleck virus. The
aphid infestation in young plants seriously
impairs ear production, while severe outbreak
at tasseling stage affects seed set. The aphid is
widely distributed and appears in serious form
on the water-stressed crop and particularly
during drought years.
M.K. Dhillon et al.
Cob borer (H. armigera): Initially it started
with infestation on Kharif maize, but currently
it damages all seasonal maize crops in Uttar
Pradesh, Bihar, Punjab, Haryana, Rajasthan,
Himachal Pradesh, Karnataka, Andhra Pradesh,
Maharashtra, and Delhi. Cob borer alone causes
up to 50 % crop loss.
Flower-eating beetle: The insect was reported
as occasional pest infesting tassel of maize in
the Hyderabad region, while in recent years, it
has also become a serious problem of maize
during tasseling stage in northern India.
Termites (Macrotermes spp. and Odontotermes
spp.): Termites have emerged as one of the
major problems in maize production across
India, including traditional maize growing
states, viz., Andhra Pradesh, Karnataka,
Bihar, and Uttar Pradesh. Termites occasion-
ally cause partial or total defoliation of maize
seedlings, but mature plants experience sever
damage. The termite infestation at the seed-
ling stage shows wilting symptoms in the
young seedlings, and complete damage to
the root system results in irreversible drying
of the plants. There is no certain pattern of
wilting; it could be on the field boundaries, in
a certain direction across the field, in patches
across the field, etc., depending on the move-
ment of the termites in the field. In older
plants, termites begin to attack the main
root system, prop roots, stems, and cobs and
eventually pack the stems with soil and cover
them with galleries or tunnels made of thin
sheets of soil. As plants mature, the amount
of damage increases rapidly and so does the
likelihood of lodging, brought about directly
by termite injury or by wind. Severely dam-
aged plants may lodge and be completely
destroyed by termites.
8.5 Management of Insect-Pests
in Maize
The maize crop is damaged by an array of insect-
pest groups with different feeding habits right
from the seedling stage to maturity of the crop,
8 InsectPests and Their Management: Current Status and Future. . .
and no single strategy is sufficient to manage
such complex group of insect-pests. Thus, here-
under are some tactics for the management of
insect-pests of maize:
Collection and burying of stubble and stalks
or plowing and destroying crop residue and
removal of infested plant parts or infested
plants through hoeing are effective in reduc-
ing the carryover populations of stem borers
and termite menace in the forthcoming crop
season.
Selection of insect-resistant maize varieties
followed by seed treatment with carbo-
furan (5 %), thiamethoxam (9 ml/kg seed),
or imidacloprid (4 g/kg seed) improves
plant stand and seedling vigor and reduces
the damage by stem borer and corn leaf
aphid.
Application of phorate 10G at 15 kg/ha in
furrows before sowing or application of 57
granules of carbofuran 3G in the central whorl
of the plants one week after germination is
effective for the management of stem borers
and shoot fly in maize.
Growing maize in association with legumes
or intercropping with soybean significantly
reduces damage by stem borers.
Conserve egg parasitoid, Trichogramma
chilonis (Ishii), and larval parasitoid,
Apanteles flavipes (Cameron), for the man-
agement of maize stem borers.
In case of heavy infestation of stem
borers, spray the crop with cypermethrin
25 EC at 0.05 % or deltamethrin 2.8 EC
at 0.02 %.
The aphid and shoot bug populations can
be effectively controlled by the cocci-
nellids [Coccinella septempunctata L. and
Menochilus sexmaculata (Fab.)] if available;
otherwise, spray the crop with monocrotophos
or dimethoate at 0.1 %.
Apply fipronil granules at 20 kg/ha
followed by light irrigation. Application of
imidacloprid or thiamethoxam drop by drop
in the water channel at the source while
irrigating the field also provides satisfactory
reduction in damage by termites.
101
8.6 Conclusion
The hybrid initiative turned out to be exceedingly
important for QPM efforts, and many countries
in the developing world, including India, are
becoming increasingly interested in QPM to
reduce malnourishment and sustainable
nutritional security. However, the yield potential
of the maize genotypes deployed in India has not
reached even half of that in the USA as a result of
different stress factors mainly the insect-pests
which cause huge crop losses. It is also likely
that the increase in concentration and quality of
nutritional compounds, particularly the protein in
QPM, might favor the proliferation of insect-
pests as they too prefer quality food for their
growth and development and could be major
constraints to increasing production and produc-
tivity of QPM. The grain yield, nutritional qual-
ity, and insect resistance are being addressed
individually; however, collective action to han-
dle all the three problems has been lacking.
Therefore, development and deployment of high-
yielding and insect-resistant QPM genotypes
under the umbrella of integrated pest management
system might help in narrowing down the yield
gap by reducing the crop losses caused by insect-
pests and could be the best fit R&D model for food
and nutritional security, environmental safety, and
sustainable maize production.
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tion methods based on evaluation-cum-recombination
block. Plant Breed Rev 14:139163
Widstrom NW, Snook ME (2001) Recurrent selection for
maysin, a compound in maize silks, antibiotic to
earworm. Plant Breed 120:357359
Williams WP, Buckley PF, Davis FM (1995) Combining
ability in maize for fall armyworm and southwestern
corn borer resistance based on a laboratory bioassay
for larval growth. Theor Appl Genet 90:275278
Williams WP, Davis FM, Buckley PF (1998) Resistance
to southwestern corn borer in corn after anthesis.
Crop Sci 38:15141517
Worku M, Banziger M, Friesen D, Erley GSA, Diallo AO,
Vivek B, Horst W (2007) Protein quantity and
quality, and grain yield performance of quality
protein maize and normal endosperm maize under
different levels of nitrogen. Afr Crop Sci Conf Proc
8:19051909
 Physiological Response of Maize Under
Rising Atmospheric CO2 and 9
Temperature

Anjali Anand, Sangeeta Khetarpal, and Madan Pal Singh

Abstract
The projections for future climate change may have a strong influence
on agricultural productivity. Maize, being a C4 plant, has evolved to
adapt to the atmospheric CO2 concentration with higher photosynthetic
efficiency than C3 plants. It is believed that C3 plants would gain a
competitive advantage under increasing CO2, but studies indicate that
C4 plants sometimes perform better due to improved water use effi-
ciency at the ecosystem level. C4 plant species have higher temperature
optima for growth than C3 plants. Temperatures above this range can
affect the photosynthetic machinery, thereby decreasing growth. Despite
the indication about the improvement in growth of C4 plants under
increasing CO2 levels, the contribution of other factors still remains
unclear in maize. This compilation is an attempt to highlight the factors
and processes affected by climate change in maize and the areas of
research that need to be strengthened to understand the underlying
mechanisms.

9.1 Introduction rise in CO2 and other greenhouse gases is

Rise in atmospheric CO2 with parallel increase
in temperature is posing to be a serious threat
to the farming community. During the last
12 years, the increasing rate of CO2 has been
1.9 l l 1 year 1 and is predicted to reach the
level of 570 l l 1 by the middle of this century
(IPCC 2007). A predicted consequence of this
A. Anand (*)  S. Khetarpal  M.P. Singh
Division of Plant Physiology, Indian Agricultural
Research Institute, New Delhi 110012, India
e-mail: anjuanand2003@yahoo.com
warming of the earths surface, which can
cause significant changes in crop productivity
through direct or indirect effects on crops, soils,
pests, etc. Environmental factors during crop
growth period determine the yield and quality
of the produce, which in turn result from the
physiological processes occurring during plant
growth and development. All the physiological
processes are affected by the change in the
environment, which finally limits productivity
of the crop. Elevated CO2 causes partial stoma-
tal closure, thereby decreasing leaf transpira-
tion, while at the same time the carbon
assimilation is increased (Morison 1998). The

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 105
DOI 10.1007/978-81-322-1623-0_9, # Springer India 2014
106
extent to which a species can acclimate to the
changing environment by maintaining an opti-
mum balance in enzymatic reactions of carbon
fixation thus gains prime importance. In this
chapter, various components affecting the
growth and yield of maize and other cereal
crops under elevated CO2 and temperature are
discussed.
C4 plants account for a small percentage of the
angiosperms, albeit they make an important con-
tribution to productivity as many agriculturally
important crops and grasslands are C4 in nature.
In C3 plants, the current atmospheric CO2 levels
limit photosynthetic capacity and growth. An
increase in CO2 levels increases photosynthesis
by reducing oxygenase activity of the photo-
synthetic enzyme, ribulose-1, 5-bisphosphate
carboxylase/oxygenase (Rubisco) and photorespi-
ration, if photosynthesis is not feedback-inhibited
by limited utilisation of the products of photosyn-
thesis (Bowes 1996; Drake et al. 1997). C4 plants
possess two types of cells, mesophyll and bundle
sheath cells, and possess a mechanism that
concentrates CO2 in the bundle sheath cells to
levels that have been estimated to be 38 times
more than the atmospheric CO2 concentration
(Kanai and Edwards 1999). Ribulose 1,5-
bisphosphate carboxylase/oxygenase (Rubisco), a
photosynthetic enzyme, is located in the bundle
sheath cells where the increase in the ratio of
CO2/O2 helps in reduction of photorespiration.
This led to an interpretation that an increase in
CO2 level may have insignificant effect on growth
of C4 plants (Reynolds 1996). However, there are
many reports of substantial increase in growth
of some C4 species under CO2 enrichment. A
28 % increase in growth of Panicum antidotale
(Ghannoum et al. 1997) and 2429 % in
Bouteloua gracilis (Chen et al. 1996) were
reported under double the ambient levels of CO2.
Daily average temperature of 2326  C resulted in
325 % increase in growth accompanied with
430 % enhancement of photosynthesis under
levels of CO2 that were double to that of ambient
conditions (Ziska and Bunce 1997). Further, the
magnitude of growth stimulation of C4 plants to
elevated CO2 increases with reduced water
A. Anand et al.
availability, temperature and light intensity
(Seneweera et al. 1998). The plants exhibit
reduced stomatal conductance and transpira-
tion rate, thereby increasing water use effi-
ciency (Ghannoum et al. 2000). A differential
response to increased CO2 levels was observed
in maize with studies showing no enhancement
in growth (Hunt et al. 1991) to 50 % stimulation
(Rogers and Dahlman 1993). The differences
were dependent on species, cultivars and environ-
mental factors like temperature, light intensity,
water and nutritional status of the plants (Drake
et al. 1997). However, these studies gave one clear
indicationthat maize has the potential to
respond to elevated CO2.
9.2 Elevated CO2 and
Photosynthesis in Maize
The basis of increased biomass production was
studied in maize plants grown in temperature-
controlled conditions under high light and CO2
(1,100 l l 1). The plants exhibited 20 %
increase in biomass and 23 % in leaf area under
elevated CO2. Light-saturated rates of photosyn-
thesis on leaf area basis indicated higher rates
of carbon fixation and dark respiration under
high CO2 levels. The plants grown at high CO2
levels had lower carboxylation efficiencies
(23 %) with lower stomatal densities in both
young and mature leaves. Maize plants benefitted
from increased CO2 by decrease in activity of
C4 enzymes, NADP-malate dehydrogenase
exhibiting the greatest decrease, without loss in
the rate of carbon assimilation. This decrease
could be explained by either difference in activa-
tion state of transcriptional or translational con-
trol of synthesis of specific proteins. The
activities of enzymes required for sucrose and
starch formation, fructose 1,6-bisphosphatase
and ADP-glucose pyrophosphorylase, increased
significantly under elevated CO2 condition by
8 and 36 %, respectively (Fig. 9.1). The increased
capacity to synthesise sucrose and starch and to
utilise these end products of photosynthesis to
9 Physiological Response of Maize Under Rising Atmospheric CO2 and Temperature 107

350 mL L-1 1100 mL L-1

60 a Rubisco p=0.10 9.3 Elevated Temperature and

50 Photosynthesis in Maize
40
p=0.62
30 9.3.1 Chlorophyll Fluorescence
20
10 Various components of photosynthesis have been
0 studied to elucidate the most sensitive compo-
60 b G3PDH p=0.14 nent to high temperature. The chlorophyll fluo-
50 p=0.01 rescence traits of maize leaves were similar to
40 that reported for C3 plants under high tempera-
30
ture (Law and Crafts-Brandner 1999). The effect
20
was evident in non-photochemical fluorescence
10
quenching (qN) well before it could be realised
0
c Ru5PK
in the maximum quantum yield of photosystem II
Activity, mol m2 s1

150
125 p=0.63
(Fv/Fm) (Figs. 9.2 and 9.3). Thylakoid
100
p=0.79 energisation was observed due to increased qN
75 at moderate leaf temperatures of 32.537.5  C,
50 which was associated with increased ratio of
25 ATP to ADP. The higher increase in temperature
0 (42.5  C) caused significant inhibition of photo-
d FBPase system II (PSII), which is otherwise considered
9 p=0.52
to be the most heat-sensitive component of pho-
6 tosynthesis (Heckathorn et al. 1998) (Fig. 9.3).
p=0.75

0 0.8
e ADPG-PP p=0.11
Fluorescence Quenching

30
Nonphotochemical

p=0.02
0.6
20
(rel. units)

10 28 8C
0.4

0 32.5 8C
8 DAE 30 DAE
Fig. 9.1 Activities of key enzymes of the C3 cycle,
sucrose and starch biosynthesis in maize leaves at
8 DAE (second leaves) and 30 DAE (sixth to seventh
leaves) from plants grown under ambient (350 L L 1,
closed bars) and three times ambient (1,100 L L 1, open
bars) CO2. (a) Rubisco, ribulose 1,5-bisphosphate car-
boxylase/oxygenase. (b) G3PDH, glyceraldehyde-3-
phosphate dehydrogenase. (c) Ru5PK, ribulose-5-phos-
phate kinase. (d) FBPase, fructose 1,6-bisphosphatase.
(e) ADPG-PP, ADP-glucose pyrophosphorylase (Source:
Maroco et al. 1999)
produce extra energy could explain the increased
growth of maize under CO2 enrichment (Maroco
et al. 1999).
0.2 37.5 8C
0
0 60 120 180 240 300
Time (sec)
Fig. 9.2 Effect of leaf temperature on qN of maize
leaves. An attached leaf was dark-adapted for 1 h at
28  C prior to conducting non-photochemical quenching
analysis. Subsequent measurements were made on the
same leaf tissue after increasing the leaf temperature at
1  C min 1 in the dark to 32.5  C for 1 h and then to
37.5  C for 1 h. Each curve represents the mean  SE of
three independent measurements. At 28  C, qN was the
same for plants that were dark-adapted for 1 and 3 h (data
not shown) (Source: Maroco et al. 1999)
108
100
80
Relative Fv/Fm (%)
60
40
20
0
30 35 40
Leaf Temperature (8C)
Fig. 9.3 Effect of leaf temperature on the Fv/Fm of
maize leaves. Intact leaves were dark adapted for 1 h at
28  C, Fv/Fm was measured, leaf temperature was
increased at 1  C min 1 to the indicated temperature,
and Fv/Fv was measured again. Different plants were
used for each temperature treatment, and the data points
represent the mean  SE of three independent
measurements (Source: Maroco et al. 1999)
This inhibition was not associated with decline in
photosynthesis up to 37.5  C in maize leaves
(Crafts-Brandner and Salvucci 2002). The
decrease in photosynthesis at higher temperature
could be through a limitation in the activation
state of Rubisco. In C3 plants, the increase in qN
due to heat stress is correlated with decreased
photosynthesis and gives an indication on inhibi-
tion of Calvin cycle activity (Feller et al. 1998).
9.3.2 Enzymes of C4 Pathway
Phosphoenolpyruvate carboxylase (PEP carbox-
ylase) and pyruvate phosphate dikinase (PPDK)
are the key enzymes of C4 pathway, and inhibi-
tion of either by heat stress, especially PPDK due
to its low activity (Furbank et al. 1997), may
decrease the supply of C4 acids available for
decarboxylation. This would further limit the
supply of CO2 to Rubisco. In maize, the activity
of PEP carboxylase and PPDK was not affected
by increase in leaf temperatures up to 45  C,
but activation state of PEP carboxylase was
A. Anand et al.
marginally sensitive to temperature of 40  C or
higher. In general, C4 plant species have higher
temperature optima for photosynthesis than
C3 plants due to the presence of a CO2-
concentrating mechanism (Edwards and Walker
1983). In C3 plants, an increase in the ratio of
oxygenase to carboxylase activities of Rubisco
under high temperature leads to inhibition of net
photosynthesis. Considering all these limitations,
the catalytic turnover rate of Rubisco increases
minimally with temperature. Thus, in C3 plants
photosynthesis declines at leaf temperature
above 32  C because the slight advantage in
45 catalytic turnover is negated by a decrease in
Rubisco activation state (Crafts-Brandner and
Salvucci 2000). As temperature increases, the
ratio of dissolved O2/CO2 and the specificity of
Rubisco for O2 increase (Jordan and Ogren
1984). Rubisco has low affinity for CO2 and
needs to be activated by a chloroplastic protein,
Rubisco activase, at physiological concentration
of CO2 and Mg2+ (Portis et al. 1986). Rubisco
activase, however, has been shown to be quite
sensitive to high temperature (Salvucci and
Crafts-Brandner 2004). In maize, the activation
state of Rubisco decreased at temperature
exceeding 32.5  C, with complete inactivation
at 45  C. The inactivation of Rubisco is the
primary constraint on the rate of photosynthesis
of maize leaves as leaf temperature increased
above 30  C. High temperature denatures
Rubisco activase rendering it unable to fit
correctly onto Rubisco. Consequently, inactive
Rubisco is not converted to active form (Crafts-
Brandner and Salvucci 2000). Rokka et al.
(2001) reported that high temperature induces
the formation of high molecular mass aggregates
of activase. The high level of CO2 in the meso-
phyll chloroplasts allowed for a substantial
temperature-dependent increase in the catalytic
turnover rate of activated Rubisco, thus
counteracting the effects of decreased activation
state until temperature approached 40  C. Levels
of 3-phosphoglyceric acid and ribulose-1,5-
bisphosphate decreased and increased, respec-
tively, as leaf temperature increased, consistent
with the decrease in Rubisco activation. When
leaf temperature increased gradually, Rubisco
9 Physiological Response of Maize Under Rising Atmospheric CO2 and Temperature
activation acclimated in a similar manner as
photosynthesis, and acclimation was associated
with the expression of a new activase polypeptide.
9.4 Elevated Temperature and
Grain Filling
During grain-filling stage, availability of photo-
synthetic assimilates and the ability of the grains
to utilise them for the synthesis of reserves are
two major activities. The optimum temperature
for dry matter accumulation in starch-storing
crops ranges between 20 and 30  C, and it is
very unusual because this temperature is lower
than the level of temperature on which proteins
start denaturing. The main reason for the low
temperature optimum for maximum grain yield
of cereal crops has been due to the effect of
temperature on both grain-filling rate and its
duration (Muchow 1990; Brooking 1993). High
temperature reduces grain size due to the steady
decrease in the duration of grain filling combined
with a failure of compensation by increased rate
of dry matter accumulation above a threshold
temperature.
Amylose to amylopectin ratio and reduced
grain density are two important components of
the grain yield, and changes in them due to any
environmental stress can also affect the grain
yield (Lu et al. 1993). Early stages of grain
development are sensitive to high temperature
than later ones (Tashiro and Wardlaw 1991),
but this can also have direct effects on starch
deposition (Bhullar and Jenner 1986), which is
related to a limitation in the supply of assimilates
to the grain (Wardlaw et al. 1980). Rijven (1986)
on the biochemical basis of the effect of temper-
ature identified the enzyme, soluble starch
synthase (SSS). Later studies in potato and barley
suggested that the site of damage in starch syn-
thesis is extra amyloplastic (Mohabir and John
1988) or involves sucrose synthase (MacLeod
and Duffus 1988). Keeling et al. (1993) have
also demonstrated that part of the effect of high
temperature on starch synthesis is linked with
SSS. Starch deposition in amyloplast involves
ADP-glucose pyrophosphorylase, a starch synthase
109
and branching enzyme (Emes et al. 2001; Zeeman
et al. 2010).
Glucose-1-phosphate is converted to ADP-
glucose by the enzyme AGPase, which is then
utilised by starch synthase. Various genetic and
biochemical studies have reported that AGPase is
a rate-limiting and regulatory enzyme in the
pathway for starch synthesis. Maize endosperm
mutants, shrunken-2 (sh-2) and brittle-2 (bt-2),
which were deficient in AGPase, showed defi-
ciency in starch content (Dickinson and Preiss
1969). These mutations involved the genes for
small and large subunits, respectively, of
AGPase (Tsai and Nelson 1966). Antisense inhi-
bition of AGPase showed reduction in starch
content in potato tubers (Muller-Rober et al.
1992). AGPase is allosterically activated by
3-phosphoglycerate (3PGA) and inhibited by Pi.
The ratio of 3PGA/Pi regulates the catalytic
activity of AGPase in the leaves (Preiss and
Sivak 1996). Studies using maize endosperm
mutants affecting branching enzyme (BE) and
sucrose synthase showed that there was only
minor control over flux by these enzymes
(Singletary et al. 1997). Further, studies with
the bt-2 mutant, which affects ADP-Glc
pyrophosphorylase activity, showed that there is
minimal control of flux by this enzyme. The
effects of reduced ADP-Glc pyrophosphorylase
on starch synthesis are related with a reduction in
grain-filling duration rather than in grain-filling
rate. SSS has been shown to be a major site of
control of flux through the pathway of starch
synthesis in both developing wheat and maize
grains (Keeling et al. 1994).
Multiple isoforms of starch synthase that
transfers glucosyl moiety of ADP-glucose to the
non-reducing end of the pre-existing -1,4-glucan
primer through an -1,4 glycosidic linkage
have been reported in all the plant tissues
(Ball and Morell 2003; Dian et al. 2005). In
cereal endosperm there are about five SS
(starch synthase) isoforms that have been
categorised according to conserved sequence
relationships. Four isoforms, named as SS I,
SS II, SS III and SS IV, are believed to have
unique functions during the amylopectin synthe-
sis, but their exact roles have not been identified.
110
It has been reported that granule-bound starch
synthase (GBSS), which is encoded by the waxy
(Wx) locus in cereals, is involved to elongate amy-
lose (Shure et al. 1983). The waxy mutants of maize
containing wild-type amount of starch with no amy-
lose are defective for GBSS (Nelson and Rines
1962). The enzyme GBSS binds tightly with the
starch granule and is linked with amylose synthesis
(Zeeman et al. 2010).
The level of temperature above 25  C has been
reported to affect the grain yield adversely because
the enzyme SSS is very heat sensitive. Two differ-
ent properties of SSS have been identified which
are affected by heat treatment and limit the grain
yield. Firstly, the rate of enzyme activity is
adversely affected by elevated temperature, and
this effect is reversible on returning to a lower
temperature. The effect of high temperature on
enzyme rate has been quantified using enzyme
Q10 and found to be suboptimal above 20  C tem-
perature. Secondly, due to prolonged period of
exposure to high temperature, loss in enzyme activ-
ity occurs due to thermal inactivation and is not
freely reversible. Thermal inactivation of enzyme
occurs when temperature exceeds 20  C in wheat,
but for similar response temperature more than
30  C is needed for maize SSS (Fig. 9.4).
Normally, the inherent stability or Q10
characteristics of other enzymes involved in
starch synthesis pathway are not affected by
high temperature except for branching enzyme,
which has minimal flux-control strength. Simi-
larly, SSS thermal inactivation may not be a
major problem in field-grown maize for develop-
ing grain, because temperature rarely is high
enough to cause enzyme inactivation. However,
the effect on enzyme Q10 is more physiologically
relevant, since maize SSS is operating
suboptimally as temperature exceeds 20  C and
selection of a high-temperature-tolerant form of
maize SSS will be a better approach, using
enzyme Q10 as a selection trait (Keeling et al.
1994). The transfer of this trait from the tropical
donor to other lines through backcross breeding
can be useful to produce a heat-tolerant line.
Other enzymatic proteins are also sensitive to
heat stress as it denatures the enzyme and causes a
loss of enzyme activity. For thermostability in the
A. Anand et al.
200.0
Relative activity
100.0
0.0
0 10 20 30 40 50
Temperature (8C)
Fig. 9.4 Effects of temperature on SSS activity obtained
from the endosperm of various crops. Data are presented
as a percentage of the activity obtained at 25  C. Results
are given as the mean of duplicate assays. Maize (), rice
(~), sorghum (), potato (), millet (), wheat ()
(Source: Keeling et al. 1994)
protein, chaperones, a specific class of proteins,
have been known for assisting other proteins in
post-translational folding and maintaining them in
a proper functional state (Ellis 1990). Heat-shock
proteins (HSPs) are well-known to work as molec-
ular chaperones; they aid in refolding proteins
denatured by heat and prevent them from
aggregating and play a very significant role in
improving tolerance to high temperature and
other stresses. Among the various classes of
HSPs, LMW-HSPs represent a set of homologous
proteins in the range of 1530 kDa (DeRocher and
Vierling 1994). During stress conditions, LMW-
HSPs may accumulate up to 1 % of the cellular
protein (Vierling 1991). They are involved to
protect the proteins associated with respiratory
electron transport in mitochondria and PS II
electron transport in thylakoid membrane of the
chloroplast. A 22 kDa HSP in the chloroplasts of
Chenopodium album has been shown to interact
with the thermolabile oxygen-evolving complex
of PS II and protect it from heat stress damage
(Sun and Montagu 2002).
These reports suggest that transfer of thermally
stable forms of these enzymes to cereals can
enhance their starch-synthesising ability and
9 Physiological Response of Maize Under Rising Atmospheric CO2 and Temperature
also stabilise the grain yield. Apart from above,
there are some transgenic approaches being cur-
rently used to stabilise the starch biosynthesis in
the following ways: (a) insertion of a gene or addi-
tional copy or copies of the gene of the wild-type
enzyme with enhanced activity, (b) modification of
the enzyme by protein engineering to achieve
alterations in the kinetic and/or allosteric behaviour
of the enzyme reaction in order to achieve thermal
stability and (c) modification of the promoter
sequences to achieve enzyme over-expression.
9.5 Elevated Temperature and
Grain Quality
Temperature plays a crucial role in determining
the quality of cereal crops. High temperature
during the grain-filling stage results in smaller
grains. While the quality of cereal crops like
wheat and rice has been extensively studied
under normal environment, there are no reports
available on the effect of changing climate on
quality of maize. Processing industries require
grains possessing specific quality attributes. The
important quality parameters in wheat are grain
hardiness, grain size, milling, dough strength,
and protein and starch quality. These attributes
are genotype-specific and also depend upon its
interaction with climate, growing temperature
and soil fertility. Grain protein and oil content
are used as grading factors in cereal trading.
The kernel size, composition and protein con-
centration are governed by post-anthesis envi-
ronmental conditions. Over 80 % of nitrogen in
the crop is taken up before flowering and stored
in stems and leaves. It is deposited in the grain in
the form of protein at a rate that reaches its
maximum during the early part of grain filling.
Carbohydrate in the form of starch is
accumulated in the grain, but its rate reaches a
maximum later during the grain-filling process.
In general, grain nitrogen concentration increases
with increase in temperature and reduced rain-
fall. In Australian variety trials extending over
27 years, grain nitrogen concentration was posi-
tively associated with the number of hours above
35  C during grain filling (Blumenthal et al. 1991).
111
In China, grain nitrogen concentration in durum
wheat was found to be positively associated with
average daily temperature during grain filling and
negatively correlated with growth period (He et al.
1990). Under controlled environmental conditions,
Ahmad et al. (1989) have reported similar relation-
ship in experiments conducted at Lahore, Pakistan.
Increasing temperature can reduce carbohydrate
accumulation more than nitrogen accumulation as
the rate of senescence is increased, thereby reduc-
ing photosynthesis and grain growth. The rate of
protein synthesis in the grain is also promoted by
warmth more than the rate of starch synthesis such
that protein concentration increases (Spiertz 1977).
9.6 Elevated CO2 and Grain Quality
There is scarcity of information on the effects of
rising atmospheric CO2 on nutritional value and
grain quality. In wheat, elevated CO2 reduces the
protein content of grain and flour by 913 %
(Rogers 1996). Plants grown under elevated
CO2 in field, open top chambers and FACE
resulted in decreased protein concentration
under elevated conditions (Fig. 9.5). Changes in
protein concentration affect dough strength and
bread making, thus being a major determinant of
grain prices (Lawlor and Mitchell 2001). The
total gluten concentration as well as
concentrations of dry and wet gluten also
decreased under high CO2 (Bencze et al. 2004).
Elevated CO2 has been reported in various
studies to cause alteration in microelements.
Concentration of all microelements decreases
up to 18.3 %, whereas macroelements are not
affected under elevated CO2 (Hogy and
Fangmeier 2008). Quality of wheat flour is
determined by a proper balance of protein and
non-protein components of grain. Poor dough of
lower extendibility and decreased loaf volume
was found in grains grown under high CO2
levels (Blumenthal et al. 1996) without affect-
ing the physiochemical properties of wheat
starch during grain filling (Tester et al. 1995).
Grain quality is a function of the sum of the
contributions of grain constituents such as
proteins, starch and lipids to dough strength,
112 A. Anand et al.

Protein GC (3)a pots < 101

GH (10)a
CTC (0)
OTC (21)b
FACE (0)

GC (1)R pots > 101

GH (0)
CTC (0)
OTC (3)R
FACE (0)

GC (0) field soil

GH (0)
CTC (11)a
OTC (14)a
FACE (2)a

20 15 10 5 0 5 10 15
Relative CO2 effect [%]

Fig. 9.5 Relative average changes ( SE) in protein
concentration of wheat grains due to CO2 enrichment
(550 vs. 380 mol mol 1) in regard to exposure system
and rooting volume. GC growth chamber, GH glasshouse/
greenhouse, CTC closed field chamber, OTC open top
chamber, FACE free-air CO2 field enrichment. Significant
CO2 effects are denoted by ***(p<0.01), **(p<0.01) and
*(p  0.05). Number in parenthesis indicates number of
studies included (k); letter indicates significant
differences between exposure systems (Source: Hogy
and Fangmeier 2008)

loaf volume and also interaction between these
components.
Oil deposition starts late during seed develop-
ment and decreases with high temperature expo-
sure resulting in decreased oil content. Fatty acid
composition also gets influenced by production
of saturated fatty acids under warmer conditions
and polyunsaturated fatty acids under cooler
moist conditions (Seiler 1983). High temperature
increases the percentage of monounsaturated
fatty acid (oleic acid) and decreases the polyun-
saturated fatty acid (linolenic acid) in mustard
(Triboi-Blondel and Renard 1999) and soybean
(Rennie and Tanner 1989). Moreover, it affects
protein content more than oil content, but oil
concentration shows an inverse relationship
with protein content. During reproductive period
of the crop under Mediterranean-type environ-
ment, adverse effects of high temperature on
oil content have been reported in canola by
Gunasekera et al. (2006). In maize, such studies
are lacking, and the quantification of increasing
environmental temperature on quality
parameters will be useful in planning agronomic
interventions and in developing ideal plant
type for future climatic scenarios. We need to
urgently take steps to increase adaptive capacity
by shifting our priority of research towards
adaptive strategies accompanied by change in
policies by the government.
9.7 Conclusion
The information on the response of maize and
other C3 crops to elevated CO2 and temperature
compiled above confirms that rising atmospheric
CO2 is beneficial for the growth and yield of C3
crops compared to C4; however, increasing tem-
perature may negate these responses particularly
during reproductive growth. On the contrary,
elevated CO2 may affect the nutritional quality
of tissues as well as grains due to its dilution
effect. Looking at agricultural perspective, rising
9 Physiological Response of Maize Under Rising Atmospheric CO2 and Temperature
CO2 may lead to reassessing the fertiliser manage-
ment, as critical nitrogen concentrations in the
tissues decrease. Since grain quality is negatively
affected by CO2 enrichment, we need to develop
genotypes with efficient nitrogen uptake for high-
CO2 environment. Concurrent increase in air tem-
perature may affect the quantity as well as quality
of grains. C4 crops are well-adapted to high
temperature environment, but extreme weather
phenomenon, as projected under changing cli-
mate, may affect their grain quality and yield.
Hence, under changing climate, identifying the
genotypes that can tolerate extreme weather
events and introgression of these traits in breeding
programmes can contribute to protect the seed
yield and quality of maize and other cereal crops
in future climate change scenarios.
References
Ahmad S, Ahmad N, Ahmad R, Hamid M (1989) Effect
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 Part V
Value Addition in Maize
 Maize Utilisation in Food
Bioprocessing: An Overview 10
Seema Sangwan, Sandeep Kumar, and Sneh Goyal

Abstract
Fermented foods are considered as palatable and wholesome foods
prepared from raw or heated raw materials and appreciated for their
attributes such as pleasant flavour, aroma, texture, improved processing
properties and better digestibility. Fermentation is carried out by the
action of a single or a group of micro-organisms including homo- and
heterofermentative lactic acid bacteria; moulds such as Mucor, Rhizopus,
Trichoderma, Aspergillus and Penicillium; and yeasts such as Saccharo-
myces cerevisiae, Schizosaccharomyces pombe, Hansenula anomala and
Debaryomyces hansenii. Globally, cereals rank number one as food crops
as well as fermentation substrates. Among cereal crops, maize with annual
worldwide production of 876.8 million metric tons during 20112012 has
been utilised in the fermented form for hundreds of years as a stimulant, in
traditional medicine, as well as in religious ceremonies. Maize is a good
source of dietary fibre and protein, while being very low in fat and sodium.
The endosperm consists of 7273 % starch embedded in a protein matrix
that makes the maize an excellent substrate for fermentation. Maize is
processed, fermented and consumed in various ways. It is usually ground
and pounded followed by boiling, baking or frying. Alternatively, the
whole grain may be boiled or roasted prior to fermentation. Maize meal
can be cooked with water either to provide a thick mush/dough or to provide
gruel, porridge or soup. In this chapter, we elaborate the processing of
maize for fermentation and illustrate the diversity, importance and
microbiological features of some fermented maize products.

S. Sangwan (*)  S. Goyal
Department of Microbiology, CCS HAU, Hisar, India
e-mail: seema_sangwan80@yahoo.co.in
S. Kumar
Germplasm Evaluation Division, National Bureau
of Plant Genetic Resources, New Delhi, India
10.1 Introduction
Food products owe their production and
characteristics to the fermentative activities of
micro-organisms in order to bring a desirable
change. Various raw materials have been

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 119
DOI 10.1007/978-81-322-1623-0_10, # Springer India 2014
120
subjected to the action of micro-organisms or
enzymes in food fermentation, thereby
converting carbohydrates into alcohol, carbon
dioxide and organic acids (William and Dennis
2011). Fermentation is the slow decomposition
process of organic substances induced by
micro-organisms, or by complex nitrogenous
substances (enzymes) of plant or animal origin
(Walker 1988). It can be described as a biochem-
ical change, which is brought about by the
anaerobic or partially anaerobic oxidation of
carbohydrates by either microorganisms or
enzymes. This is distinct from putrefaction,
which is the degradation of protein materials
(Battcock and Ali 1998). Fermented foods
originated many thousands of years ago when
presumably micro-organisms contaminated local
foods, but nowadays, these foods comprise about
one-third of the worldwide consumption of food
and 2040 % (by weight) of individual diets.
Globally, cereals rank number one as food crops
as well as fermentation substrates. Traditional
fermented foods prepared from most common
type of cereals, such as, rice, wheat, maize, millet
and sorghum, are well known in many parts of the
world, such as in Africa. Some are utilised as
colourants, spices, beverages and breakfasts or
light meal foods, while a few of them are used as
main foods in the diet. Among cereal crops, maize,
which has its origin in Mexico, has been consumed
in the fermented form for hundreds of years.
A number of traditional products based on maize
were developed by the indigenous populations of
Mexico and Peru in the pre-Columbian era and
utilised as stimulants, in traditional medicine, as
well as in religious ceremonies.
Maize is third behind wheat and rice, in terms
of total area under cultivation in the world. About
two-thirds of this area is in developing countries,
where maize is widely grown for direct human
consumption. As reported by Foreign Agricul-
tural Service/USDA, total annual worldwide
production of maize was 876.8 million metric
tons in the year 20112012 showing 5.54 %
increase over the previous years production.
The United States is the leading producer of
maize in the world yielding about 35.8 % of the
total or 313.9 million metric tons; China and
S. Sangwan et al.
Brazil follow on the second and third positions,
respectively. India positioned at seventh place,
after Mexico, having 2.4 % of worlds total pro-
duction or 21 million metric tons. Maize spread
to the rest of the world due to its ability to grow
in diverse climates. Sugar-rich varieties called
sweet corn are usually grown for human con-
sumption, while field corn varieties are used for
animal feed and as chemical feedstock.
10.2 Common Uses of Maize
Maize can be used for many purposes as every
part of the plant has commercial value. It is wet-
milled to separate the grain into components like
starch, oil, protein and fibre, which are then
processed into multitude of food and industrial
products including starch, sweeteners, corn oils,
beverages and industrial alcohols as well as fuel
alcohols. New bioproducts such as amino acids,
antibiotics and degradable plastics are increasingly
being synthesised using maize as a raw material.
Maize is utilised in making cornbread, by mixing
the meal with wheat flour. Immature cobs, prefer-
ably sweet corn, are boiled and eaten, while more
mature cobs are roasted. Cornstarch (maize flour)
is a major ingredient in home cooking and in many
industrialised food products. It can be hydrolysed
and enzymatically treated to produce syrups,
particularly high-fructose corn syrup, a sweetener.
Alternatively, hydrolysed cornstarch may be
fermented and distilled to produce grain alcohol.
Grain alcohol from maize is traditionally the
source of Bourbon whiskey. Starch from maize
can also be made into plastics, fabrics, adhesives
and many other chemical products.
Further, maize has carotene, a type of
carotenoids, otherwise occurring mainly in plants
and algae. Carotene can be converted into
vitamin A (retinol) by normal metabolic processes
in human beings. Vitamin A is very important to
human health, but most especially for vision, and
as an antioxidant.
In the United States and Canada, maize grains
are used as feed for livestock; however, the
digestibility and palatability of maize silage,
made by fermentation of chopped green corn
10 Maize Utilisation in Food Bioprocessing: An Overview
Table 10.1 Major benefits of fermentation
Effects of fermentation
Conditioning by steeping for wet milling of grains
Affecting sensory properties such as aroma, taste, colour
and texture
Generation of reducing sugars by saccharification prior to
alcoholic fermentation
Preservation which relies mainly on acidification and or alcohol
production
Lactic acid bacteria (LAB) isolated from fermented foods
displayed probiotic properties such as hypolipidemic,
hepatoprotective and antibacterial
Enhancing food safety by inhibition of pathogens, such as
Burkholderia gladioli which is responsible for bongkrek
poisoning in products made from presoaked corn
Removing anti-nutritional compounds such as phytate, enzyme
inhibitors, polyphenols and cyanogenic compounds
Increase bioavailability of components by affecting
physico-chemical properties of starch and associations of
fibre constituents with vitamins, minerals or proteins
Removing undesirable compounds such as mycotoxins,
endogenous toxins, cyanogenic compounds and flatulence-
producing carbohydrates
stalks, is greater than dried. Maize meal is also a
significant ingredient of some commercial ani-
mal food products, such as dog food.
Stigmas from female maize flowers, popularly
called corn silk, are sold as herbal supplements.
The corn steep liquor, a plentiful watery by-
product of maize wet-milling process, is widely
used in the biochemical industry and research as a
culture medium to grow many kinds of micro-
organisms. Some forms of the plant are occasion-
ally grown for ornamental use in the garden.
Maize cobs are also increasingly used as a
substrate for biofuel production. Ethanol produced
from maize can be mixed with gasoline to
decrease the amount of pollutants emitted when
used to fuel motor vehicles. Maize is widely used
in Germany as a feedstock for biogas plants.
10.3 Benefits of Fermentation
Fermented foods can, in general, be described as
palatable and wholesome foods prepared from
raw or heated raw materials. They are generally
appreciated for attributes such as pleasant fla-
vour, aroma, texture and improved cooking and
121
Reference
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Namugumya (2009)
Tang et al. (2011), Godliving and Mtui (2012)
Auerbach et al. (2000), Driehuis et al. (2001), Owen
(2002), Danner et al. (2003)
Oyetayo and Osho (2004), Aderiye et al. (2007)
Motarjemi and Asante (2002), Ejigui et al. (2005)
Awada et al. (2005), Ejigui et al. (2005), Mbata et al.
(2009)
Awada et al. (2005), Chavan and Kadam (1989)
Adegoke et al. (1994), DSouza and Brackett (1998),
Westby et al. (1997), Oluwafemi and Ikeowa (2005)
processing properties (Holzapfel 2002). In gen-
eral, during the production process, the fermen-
tation steps aim to achieve the properties detailed
in Table 10.1.
10.4 Microbiology of Fermented
Foods
Traditionally, the fermenting organisms came
from the natural microflora or a portion of the
previous fermentation and it is termed as starter
culture. In many cases, the natural microflora
is inefficient, uncontrollable and unpredictable
or is destroyed during preprocessing stages such
as pasteurisation. It is always beneficial to use
previously characterised, well-identified starter
culture (Table 10.2) as it can provide particular
characteristics to the food in a more controlled
and predictable manner.
The most important fungi involved in industrial
fermentation are from two of the main classifica-
tion groups: the aseptate zygomycota, which
includes Mucor and Rhizopus and the septate
deuteromycotina (fungi imperfecti), which
includes the genera Trichoderma, Aspergillus,
122 S. Sangwan et al.

Table 10.2 Commonly used starter cultures in fermentation

Microorganisms Examples Product
Yeast Saccharomyces cerevisiae, S. rouxii, S. ellipsoideus 2 Ethanol + 2 CO2 per hexose
molecule
Homofermentative Pediococcus, Lactococcus lactis, Streptococcus thermophilus, 2ATP + 2Lactate per glucose
LAB Vagococcus, some Lactobacilli molecule
Heterofermentative Carnobacterium, Oenococcus, Enterococcus, Lactosphaera, 1ATP + 1Lactate + 1
LAB Weissells, Leuconostoc mesenteroides subspecies cremoris, Ethanol + 1 CO2 per glucose
Leuconostoc mesenteroides subspecies lactis, some Lactobacilli molecule
Moulds Aspergillus oryzae, Erwinia dissolvens, Rhizopus oligosporus Variable

Penicillium, Aureobasidium and Fusarium. Yeasts
are unicellular fungi that generally reproduce by
budding; however, some exceptional species
reproduce by binary fission such as Schizosac-
charomyces pombe. Saccharomyces is the most
widely used yeast in industrial fermentations and
has applications in alcohol production and baking.
All the strains ferment glucose and many ferment
other plant-associated carbohydrates such as
sucrose, maltose and raffinose but none can
ferment lactose. Kluyveromyces lactis, which
contains the necessary lactose-transporting and
degrading enzymes, is particularly useful in
production of alcohol and biomass from whey.
Zygosaccharomyces rouxii is specially associated
with fermentation of plant products at high salt
concentration and low water activity. Many strains
like Hansenula anomala and Debaryomyces
hansenii can also grow in fairly concentrated salt
solutions and the latter is frequently isolated from
brined meat products and fermented sausages.
Lactic starters constitute the major group of
fermentative organisms that includes bacteria
having ability to convert sugars to lactic acid,
for example, Lactococcus lactis. This group
comprises 11 genera of gram-positive bacteria,
that is, Carnobacterium, Oenococcus, Entero-
coccus, Pediococcus, Lactococcus, Streptococ-
cus, Lactobacillus, Vagococcus, Lactosphaera,
Weissells and Leuconostoc. The lactic acid bac-
teria are largely mesophilic and generally grow
over a temperature range of about 1040  C,
having a temperature optima between 25  C and
35  C. Alternatively, Lactobacillus delbrueckii
subspecies bulgaricus is thermoduric and can
grow above 40  C. Similarly, the optimum pH
range varies from 4 to 8, though, some can grow
at a pH as low as 3.2 and as high as 9.6. Heterofer-
mentative organisms like Leuconostoc citrovorum
and Leuconostoc dextranicum are particularly
desirable for development of flavour and aroma
compounds such as acetyl aldehyde and diacetyl.
A good starter culture will convert most of the
sugars to lactic acid, increase the lactic acid con-
centration to 0.81.2 % (titratable acidity) and drop
the pH to 4.34.5.
10.5 Biochemistry of Fermented
Foods
Structurally, maize consists of an embryo (germ)
and an endosperm enclosed by an epidermis and
a seed coat (husk). While the germ is basically a
package of nutrients such as amino acids, sugars,
lipids, minerals, vitamins and enzymes, the husk
is comprised mainly of cellulose, pentosans,
pectins and minerals. Depending on the variety,
maize may contain a number of important B
vitamins, folic acid, vitamin C and precursor to
vitamin A. Maize is also rich in phosphorus,
magnesium, manganese, zinc, copper, iron and
selenium and has small amounts of potassium
and calcium. Maize is a good source of dietary
fibre and protein, while being very low in fat and
sodium. However, maize is naturally deficient in
lysine and tryptophan, which are two of the eight
amino acids regarded as essential for humans, but
Quality Protein Maize (QPM) has been bred to be
high in lysine and tryptophan. Maize lacks the
10 Maize Utilisation in Food Bioprocessing: An Overview
protein gluten of wheat and, therefore, makes
baked goods with poor rising capability. The
endosperm consists of starch granules of differ-
ent sizes embedded in a protein matrix, which
makes the maize an excellent substrate for
fermentation. Starch constitutes, approximately,
7273 % of the kernel weight, whereas other
carbohydrates are simple sugars that present as
glucose, sucrose and fructose in amounts that
vary from 1 % to 3 % only. Chemically, this
starch is a complex heterogeneous biopolymer
composed of amylose and amylopectin, two
high molecular weight components that may be
present in different ratios. The former is a linear
polymer containing 702,100 glucose units
linked via -1,4 glucosidic linkages, whereas
the latter is a branched polymer with 46 %
-1,6 glucosidic linkages at branched points;
the average length of branch chain is 2025
glucose units (Stewart and Russell 1987).
Commonly used starter cultures of yeast and
lactic acid bacteria lack the ability to transform
this substrate to simple sugars. Clearly, it would
require a hydrolysis to glucose by amylases, prior
to fermentation. Western techniques use endoge-
nous starch-degrading enzymes produced in the
grains through the process of malting. In malting,
the grain is moistened by steeping in water and
is allowed to germinate. During germination,
hydrolytic enzymes present in the aleurone layer
surrounding the grain endosperm attack the
endosperm, mobilising the nutrients and energy
reserve, starch. Some native cultures in South
America use salivary amylase for starch hydro-
lysis by simply chewing the substrate, while
some others utilise enzymes produced by co-
cultured, resident, or externally added moulds.
These techniques, however, are not amenable at
large-scale industrial production. Therefore,
amylolytic enzymes, a group of starch-splitting
enzymes, are of considerable importance to fer-
mentation of grains like maize at industrial level.
Starch cannot be converted to sugars easily, as it
requires prior gelatinisation by heat treatment,
liquefaction by -amylase and saccharification
by amyloglucosidase. The enzyme-mediated
hydrolysis of starch is a rapid process with little
contamination by reversion products and forma-
tion of fewer by-products. In addition, it is more
123
specific and higher yields of sugars as well as
alcohol and lactic acid are obtained.
Carbon, nitrogen and other nutrients from
surroundings of micro-organisms enter the cell
and transform into either new cell material or the
product, through a process called metabolism.
These transformations require energy, and since
most of the micro-organisms involved in indus-
trial fermentations are heterotrophs, therefore,
this energy is obtained from breakdown of
organic compounds. While in aerobic or respira-
tory processes, organisms are able to completely
oxidise substrate into CO2 + H2O, resulting in
maximum energy production, in anaerobic or
fermentative metabolism, cells are less efficient
in conversion of organic substrate into cellular
material and usually excrete partially degraded
intermediates, yielding lesser energy.
Inside the cell, the sugars are broken down by
one of the three pathways: the Embden Meyerhof
Parnas (EMP) pathway, hexose monophosphate
(HMP) pathway and Entner Doudoroff (ED)
pathway. The EMP pathway most widely occurrs
in animals, plants and fungal, yeast and bacterial
cells for glucose utilisation. A few bacteria
including pseudomonas species, which do not
metabolise glucose via EMP pathway, utilise
ED pathway. HMP pathway is especially useful
in generating precursors of aromatic amino acids
and vitamins and the supply of NADPH and H+
needed for many biosynthetic pathways. A sig-
nificant product for all the three pathways
mentioned earlier is pyruvic acid, which is
channelled into tricarboxylic acid (TCA) cycle
in aerobic metabolism and in the precursor for
various acids, alcohols and other end products in
anaerobic metabolism.
Lactic acid bacteria are gram-positive, non-
spore-forming rods or cocci, mostly aero-tolerant
anaerobes that lack cytochromes and porphyrins
and are, therefore, catalase- and oxidase-negative.
Some do take oxygen through the mediation of
flavoprotein oxidase, which is used to produce
hydrogen peroxide and/or to reoxidise NADH
produced during dehydrogenation of sugars.
Cellular energy is derived from fermentation of
carbohydrates to produce peripherally lactic acid
using two different pathways: homofermentative
and heterofermentative (Fig. 10.1).
124 S. Sangwan et al.

Homofermentatative LAB Hetrofermentatative LAB

Glucose ATP
2 ATP ADP
2 ADP Glucose-6 Phosphate

Fructose 1,6-bisphosphate NAD+

NADH
6 Phospo gluconate

NAD+
Aldolase
NADH

Ribulose-5 Phosphate + CO2

Pentoses
2 Glyceraldehyde Dihydroxy
3-Phosphate Acetone Phosphate Xylulose-5 Phosphate

Pi Phosphoketolase
Pi
+
NAD
NADH Glyceraldehyde 3-Phosphate

NAD+
NADH
2(1,3-bisphosphoglyceric acid) Pi
1,3-bisphospho Acetyl-Phosphate
glyceric acid
2 ADP NADH
2 ATP 2ADP
NAD+
2ATP

2 Pyruvate- Acetaldehyde
Pyruvate-
NADH NADH
NADH
NAD+ NAD+
NAD+
2 Lactate-
Ethanol
lactate-

NET Gain = 1ATP + 1Lactate + 1

NET Gain = 2ATP + 2Lactate Ethanol + 1 CO2

Fig. 10.1 Lactic acid fermentation using homofermentative and heterofermentative bacteria

Homofermentative lactic acid bacteria level phosphorylation. NADH produced during

contain aldolase, a key enzyme of glycolysis,
and produce virtually a single product, that is,
two molecules of lactate per glucose molecules.
They follow the Embden Mayerhoffs Parnas
(EMP) pathway where the six-carbon molecule
glucose is phosphorylated and isomerised before
cleavage by enzyme aldolase into glyceraldehyde-
3-phosphate and dihydroxyacetone phosphate.
These intermediates can be further converted
into two pyruvate and finally two lactate
molecules, generating 2 ATP by substrate-
the oxidation of glyceraldehyde-3-phosphate to
1,3-bisphosphoglycerate is reoxidized to NAD+
in the formation of lactate from pyruvate by
the action of lactate dehydrogenases (LDHs).
Heterofermentative LAB lacks aldolase and
thus cannot break down fructose bisphosphate to
triose phosphate. Instead, they oxidise glucose-6-
phosphate to 6-phosphogluconate and then
decarboxylate this to pentose phosphate, which
is then converted into triose phosphate and acetyl
phosphate by key enzyme phosphoketolase. In
10 Maize Utilisation in Food Bioprocessing: An Overview
heterofermentative LAB, triose phosphate is
converted ultimately to lactic acid with the produc-
tion of one ATP molecule, whereas, to achieve
redox balance, acetyl phosphate is reduced by
NADH to ethanol, without the generation of
any ATP. These heterofermentative bacteria can
receive additional ATP molecules through conver-
sion of acetyl phosphate to acetate.
10.6 Fermented Products of Maize
Maize is prepared and consumed in various
ways. It is usually ground and pounded. The
meal may be boiled, baked or fried. The whole
grain may be boiled or roasted and it may be
fermented. Maize meal can be cooked with
water to provide a thick mush or dough. It may
be cooked with water to provide gruel, porridge
or soup. In this section, we illustrate the diver-
sity, importance and microbiological features of
some fermented maize products (Table 10.3).
10.6.1 Chicha
Chicha is a clear, yellowish, effervescent, alco-
holic beverage prepared from maize, having
alcohol content that varies between 2 % and
12 % (v/v), most popular in South and Central
America. It has a flavour similar to that of cider
and colour varies from red to purple depending
upon the pigmented maize varieties used for its
preparation.
The traditional production of chicha is a
unique fermentation process where saliva serves
as the source of amylase for converting starch to
fermentable sugars. Alternatively, malting (ger-
mination) of maize kernels to produce the amy-
lase required for starch conversion is widely used
in modern-day processing. Frequently, a combi-
nation of both the processes can be used for
malting to yield chicha (Steinkraus 1996).
For starch hydrolysis, dry-grounded maize is
slightly moistened with water. The maize dough
thus obtained is rolled into a ball, popped into the
mouth and thoroughly mixed with saliva using
the tongue to yield gob, which is then flattened
125
against the roof of the mouth with the tongue and
placed under the sun to dry. The dried product,
termed muko, is then subjected to grinding in a
stone mill. Muko production is generally carried
out as a social event by groups of older women,
sometimes with the help of young girls. In an
alternative method the overnight soaked, maize
kernels kept in the dark for about 34 days, under
moist conditions for germination. With emer-
gence of plumules of 0.250.5 cm in length, the
kernels are heaped and covered with burlap for
12 days and the temperature is allowed to rise
until it is uncomfortable to place the hand in the
mass of kernels. The kernels become white,
parched and covered with a thin layer of ash.
Germinated grains are sun-dried, following
which they are finely ground in a stone mill.
Methods of extracting the maize flour vary
rather widely. In one of the methods, working
capacity of an earthenware pot is filled with 1:2
ratio of maize flour and cold water and heated to
approximately 75  C, followed by thorough
mixing for 1 h before allowing the mixture to
settle and cool down. When muko is used, addi-
tional non-salivated flour may be added along
with crude sugar or squash pulp. Three layers
are formed during settlement: a top liquid layer
called upi, a middle jelly-like layer and a bottom
layer that contains coarse particles (hanchi).
The upi is scooped and transferred to another
earthenware pot. The middle layer is placed in a
shallow pan, heated and concentrated to a sugar-
like product. The hanchi is pressed and filtered
and the filtrate thus obtained is added to the upi.
The upi is caramelised by simmering for several
hours and referred to as misqui kheta, which is
then allowed to cool and ferment for 46 days.
Since the fermentation pots are never cleaned,
no inoculum is required, usually for such
fermentations. After completion of fermentation,
froth is removed with a cupped hand and the
chicha is transferred to narrow-mouth pots for
consumption.
The chicha is ready for consumption when the
sweetness disappears and the flavour becomes
semi-sharp. Brown sugar or molasses may be
added in order to increase the alcoholic content
of the chicha (Steinkraus 1996).
126 S. Sangwan et al.

Table 10.3 Fermented food products from maize

Name of the
fermented Description/
food pretreatment Microorganism Nutritive value/applications Reference
Chicha Salivation and/or Saccharomyces Vitamins B Steinkraus (1996)
malting cerevisiae, S.
pastorianus,
Mycoderma vini,
Oidium lactis, Monilia
candida, Leuconostoc,
Lactobacillus,
Acetobacter,
Aspergillus,
Penicillium
Tesguino Malting and S. cerevisiae, Rich in vitamins and Ulloa et al. (1987),
extraction of Lactobacillus, enzymes & serves as a Wacher-Rodarte (1995)
juice of maize Streptococcus, refreshing alcoholic drink
stalk followed by Leuconostoc,
boiling of juice Pediococcus,
Saccharomyces,
Candida,
Cryptococcus,
Hansenula,
Brettanomyces, Pichia,
Geotrichum and
Penicillium
Umqombothi Malting Lactic acid bacteria and Vit B mild alcohol, sour Bleiberg (1979), Coetzee
yeasts aroma (1982)
Busaa Opaque maize L. helveticus, L. Crude protein, thiamine and Farnworth (2003), Willis
(Nigeria, beer salivarius, riboflavin (2002), Obot (2007), Papas
Ghana) Pediococcus et al. (2010)
damnosus, P. partulus,
Candida krusei, S.
cerevisiae
Atole Steeping and Lactic acid bacteria Sour nonalcoholic porridge, Escamilla-Hurtado et al.
(Mexico) milling of maize diacetyl provides sensory (1993)
grains properties
Pozol Boiling of kernels Leuconostoc Control of diarrhoea; Ulloa et al. (1987),
mesenteroides, antagonistic to many Canas-Urbina et al. (1993),
Lactobacillus pathogenic bacteria, yeast Nuraida et al. (1995)
plantarum, L. and moulds; acidic flavour
confusus, Lactococcus and aroma impart the
lactis, L. raffinolactis, refreshing properties
Candida krusei,
Trichosporon
cutaneum, Hansenula
fabiani, Kluveromyces
fragilis, Candida
guillermondii, C.
parapsilosis, C.
tropicalis, S.
Cerevisiae,
Geotrichum candidum,
Trichosporon
cutaneum, Mucor
rouxianus
(continued)
10 Maize Utilisation in Food Bioprocessing: An Overview 127

Table 10.3 (continued)

Name of the
fermented Description/
food pretreatment Microorganism Nutritive value/applications Reference
Ogi (Nigeria, Gelatinisation Corynebacterium Soft or stiff gel Control Nkama et al. (2000), Achi
Benin) and Hydrolysis (hydrolyses), L. gastroenteritis in animals (2005), Osungbaro (2009),
plantarum, and man, Rich in proteins, Anigo et al. (2009),
Saccharomyces vitamins and minerals Muhimbula and
cerevisiae, Candida Issa-Zacharia (2010),
mycoderma, Aderiye et al. (2007),
Rhodotorula, Adebolu et al. (2007),
Cephalosporium, 2012
Fusarium, Aspergillus,
Penicilliums
Mahewu Based on corn Streptococcus lactis, L. Nonalcoholic sour beverage Gadaga et al. (1999),
(East Africa) meal cellobiosis, L. brevis, L. Odunfa et al. (2001),
delbrueckii, L. Steinkraus (1996)
Bulgaricus, Candida
krusei, S. Cerevisiae,
Candida
guilliermondii
Pito (Nigeria Meshing of Geotrichum candidum, Dark brown alcoholic Akyeampong (1995), Sefa-
and Ghana) malted maize Lactobacillus sp., liquid, varies in taste from Dedeh (1999)
Candida sp. sweet to bitter
Uji (Sudan) Grounded and Lactobacillus Sour maize gruel Mbugua (1984)
slurrified plantarum, L.
fermentum, L.
cellolbiosus, L.
buchneri
Abati Based on maize Lactic acid bacteria Alcoholic beverage Haard et al. (1999)
(Paraguay & dough or flour and yeasts
Argentina)
Acupe Malting Lactic acid bacteria Fermented and sweetened Haard et al. (1999)
(Venezuela) and yeasts nonalcoholic beverage
Cachiri Based on maize, Lactic acid bacteria Fermented nonalcoholic Haard et al. (1999)
(Brazil) manihot or fruit and yeasts beverage
and produced in
clay pots
Agua-agria Grinding of Lactic acid bacteria Nonalcoholic beverage Haard et al. (1999)
(Mexico) maize with and yeasts
water
Atole agrio Fermented black Lactic acid bacteria Nonalcoholic porridge Haard et al. (1999)
(Mexico) maize dough and yeasts
Fuba (Brazil) Malting Lactic acid bacteria Nonalcoholic beverages Haard et al. (1999)
and yeasts
Ostoche Based on maize Lactic acid bacteria Alcoholic beverage Haard et al. (1999)
juice and brown and yeasts
sugar

10.6.2 Tesguino ethnic groups of northern and northwestern

Mexico. Methodologies used in the preparation
Tesguino is a slurry-like, alcoholic beverage of tesguino vary among ethnic groups. In a most
prepared by fermentation of germinated maize common approach, about 10 kg of dry maize
or maize stalk juice. It is consumed by several kernels are soaked in water for several days,
128
drained and placed either in baskets in the dark or
in a hole in the ground in order to allow them to
germinate. The germinating kernels are protected
from light, in order to prevent the formation of
green and bitter sprouts. The germinated kernels
are ground and boiled for about 8 h in water until
the mixture turns yellow. The liquid portion is
then transferred to a clay pot, and catalysts are
added. The most common catalysts are bark
(batari) or kakwara (Randia echinocarpa, R.
watsoni and R. laevigata) and kaya (Coutarea
pterosperma), which are chopped, ground and
boiled for many hours prior to being added to
the tesguino.
At higher altitudes where pine trees grow, the
catalysts used are leaves of Stevia serrata,
Chimaphila maculata and Datura meteloides;
stems of Bromus arizonicus; and roots of
Phaseolus metcalfei and Plumbago. The mixture
is allowed to ferment for several days prior to
consumption (Taboada et al. 1977; Steinkraus
1996).
In order to prepare tesguino from maize
stalks, the raw material either fresh or dry is
macerated by pounding with a club, in the
depression of a rock. The macerated material is
then placed on a sieve, water is slowly poured
over and juice is collected in a hollow pumpkin.
The juice is mixed with water and boiled for
several hours prior to the addition of catalysts.
The mixture is allowed to ferment in a dark place
for 23 days, until it develops a pleasant appear-
ance and flavour before it is consumed.
S. cerevisiae is an important micro-organism
in the alcoholic fermentation of tesguino and is
maintained on surfaces of the utensils and clay
pots that are used exclusively for the prepara-
tion of tesguino. The bacteria produce the lactic
and acetic acids which give tesguino some of its
distinctively refreshing, acidic flavour. Abundant
yeast species consistently identified at various
stages of the fermentation are alcohol producers
and include Candida guilliermondii, Hansenula
anomala, S. cerevisiae, and S. kluyveri. S.
cerevisiae and S. kluyveri. Other yeasts produce
oxidative esters that contribute to turbidity,
aroma and flavour. Compositionally, tesguino
contains 73.9 % moisture, 2 % protein, 0.21 %
crude fibre, 2.5 mg/100 g iron, 0.03 mg/100 g
S. Sangwan et al.
thiamine, 0.03 mg/100 g riboflavin and 0.29 mg/
100 g niacin. During the fermentation, protein
content increased by 58 % and lactic acid, acetic
acid and ethanol concentrations were 0.41 %,
0.11 % and 3.73 %, respectively. At the end of
the process, total protein concentration
approached 13.2 % (Wacher-Rodarte 1995).
10.6.3 Umqombothi
Umqombothi is popular among the South Afri-
can population. Maize and sorghum, used in
combination, are the most common cereals
used in South Africa to make umqombothi.
It is a pink, opaque, mild alcoholic drink having
yoghurt-like flavour and thin consistency. It is
consumed in the active state of fermentation and
therefore has a short shelf life of 23 days
(Coetzee 1982). Equal amount of maize meal,
crushed maize malt and crushed sorghum malt
are mixed with warm water in a cast-iron pot.
The mixture is left overnight for fermentation
that generates a sour odour and bubbling. A
small portion of the wort is removed and put to
one side and the remaining mash is cooked until
crusty sediment forms which can be eaten as
porridge. When making beer, the sediment is
left to cool for a day. After the mixture has
cooled, it is poured into a large plastic vat
along with the wort that was set aside. Some
additional sorghum and maize malt is added to
the vat and the brew is stirred properly. The vat
is put in a warm place overnight, to encourage
fermentation. Lactic acid bacteria and yeasts are
thought to be the predominant micro-organisms
during this fermentation. When the brew is ready,
the fermented mash is filtered through a large
metal strainer, to remove the spent grains.
Strained beer is poured into a large communal
drum and it is ready for consumption.
10.6.4 Busaa
Busaa is a Kenyan opaque maize beer having
24 % ethanol and 0.51 % lactic acid. It is
considered more nutritious and superior as com-
pared to clear lager beer due to its richness in
10 Maize Utilisation in Food Bioprocessing: An Overview
crude protein, thiamine and riboflavin and is
typically made from the most plentiful source
of grain, whether maize, millet or sorghum.
Although brewing and selling traditional drink
is illegal in Kenya, drinking traditional brew is
a common activity during many social and reli-
gious ceremonies. For example, drinking busaa is
integral to the custom of group genital
circumcisions, weddings and funerals (Willis
2002). Once controlled by elder men, drinking
of traditional brew has been largely transformed
into a commercial enterprise that includes female
and young consumers (Willis 2002; Papas et al.
2012) and is believed to be more frequently
brewed and sold by women today (Holtzman
2001; McCall 1996; Obot 2007).
At the maize-souring stage, the micro-organisms
involved are lactic acid bacteria and a few yeasts,
mainly representatives of L. helveticus, L. salivarius,
Pediococcus damnosus, P. partulus, Candida krusei
and S. cerevisiae. The main fermentation is also
governed by a mixture of yeasts and lactic acid
bacteria including C. krusei and L. casei var.
rhamnosus.
10.6.5 Atole
Atole is a sour porridge-type product prepared
from maize, in Southern Mexico. It is produced
by steeping maize grains in water for 4 days,
followed by milling during which lactic acid
fermentation takes place for 1 day. Lactic acid
bacteria present in the atole form diacetyl, which
contributes to the characteristic sensory
properties of the product.
10.6.6 Pozol
Pozol is fermented maize dough formed into
balls of various shapes and sizes. It is advanta-
geous as it can be preserved without refrigeration
under the tropical conditions, in which it is
routinely eaten, owing to its low pH, along with
having improved nutritional quality such as high
protein, niacin, riboflavin, lysine and tryptophan,
129
due to the development of certain bacteria, yeasts
and moulds.
In the production process of pozol, 11.5 kg
of kernels are boiled for 1 h in a pot containing
12 l of an approximately 10 % (w/v) calcium
hydroxide solution. During boiling, the majority
of micro-organisms associated with maize kernels
are destroyed and swelling of the kernels takes
place, thus allowing the pericarp to be relatively
easily peeled off the kernels. The kernels are
cooled, rinsed with water and termed as
nixtamal. The nixtamal is ground to obtain coarse
dough, which is manually shaped into balls. The
balls are fermented for 114 days after wrapping
in banana leaves to prevent desiccation. Ground
cacao beans or coconut may be added to the
dough prior to fermentation, to yield a fermented
product called chorote (Canas-Urbina et al.
1993). During the initial 24 h (pH 7.3) of pozol
fermentation, lactic acid bacteria (104106/g)
outnumber yeasts (102104/g) and moulds
(<103/g) and are probably responsible for the
majority of acid produced. After incubation for
30 h at 28  C, the counts increase to 1010/g for
lactic acid bacteria, 106/g for yeast and 104/g for
moulds, while the pH decreases to 4.6 (Wacher
et al. 1993).
Pozol is consumed in the southeastern states
of Mexico, such as Chiapas, Tabasco, Campeche
and Yucatan, and on a smaller scale in Veracruz,
Oaxaca and Guatemala (Ulloa et al. 1987). Pozol
balls are diluted with water in 1:21:3 ratios at
various stages of the fermentation process to
produce a whitish porridge that is consumed in
the uncooked state as a basic food in the daily
diet of large communities. Salt, toasted ground
chilli pods, sugar or honey may be added as per
the taste of consumers.
10.6.7 Ogi
Ogi porridge or sour gruel is obtained as a result
of the submerged fermentation of some cereals
such as maize in West Africa and sorghum and
millet in the North. Ogi is usually a cooked gel of
variable degree of stiffness and often marketed as
a wet cake wrapped in leaves or transparent
130
polythene bags. Gelatinised ogi is an important
indigenous, traditional weaning food common in
the whole of West Africa.
Ogi is an example of traditional fermented food,
involving a simple technique such as malting and
fermentation, which has now been upgraded to a
semi-industrial scale (Achi 2005). Microbiological
and nutritional studies by Akinrele (1970) showed
that the lactic acid bacterium Lactobacillus
plantarum, the aerobic bacteria Corynebacterium
and Aerobacter, the yeasts Candida mycoderma,
Saccharomyces cerevisiae and Rhodotorula and
moulds Cephalosporium, Fusarium, Aspergillus
and Penicillium are the major organisms responsi-
ble for the fermentation and nutritional improve-
ment of ogi. Corynebacterium hydrolyses the corn
starch but the predominant starter in the fer-
mentation production of lactic acid is L. plantarum.
Saccharomyces cerevisiae and Candida
mycoderma contribute to the flavour development
(Odunfa and Adeyele 1995).
The production of ogi is often undertaken by
unskilled female attendants of small-scale enter-
prise (Aminigo and Akingbala 2004). Therefore,
these slurries have been known to exhibit health-
promoting properties such as control of gastroen-
teritis in animals and man. This has been
demonstrated by locals who drink the raw slurry
when there is a case of diarrhoea (Oyetayo and
Osho 2004; Aderiye et al. 2007; Adebolu et al.
2012). Ogi liquors prepared using cold and hot
water methods were effective in inhibiting the
growth of most of the test organisms except
Enterobacter sp and Staphylococcus aureus;
the growth inhibition, however, was not as wide
as that of some of the antibiotics such as cipro-
floxacin, gentamicin and ofloxacin but in most
cases superior to that of tetracycline,
nitrofurantoin and cotrimoxazole (Adebolu
et al. 2007).
DogiK, an improved ogi formulated to aid the
control of diarrhoeal diseases, was produced by
using Lactobacillus starter cultures with antimi-
crobial activity against diarrhoeagenic bacteria
and also possessing amylolytic activity. The sur-
vival of diarrhoeagenic bacteria was investigated
in locally fermented ogi and in DogiK. The
foods were inoculated with cell suspensions of
S. Sangwan et al.
Salmonella, Shigella, Campylobacter,
Aeromonas, Pleisiomonas, Enteropathogenic
and Enterotoxigenic Escherichia coli, Yersinia
enterocolitica and Vibrio cholerae. None of the
diarrhoeagenic bacteria were detected in DogiK
after 6 h, whereas in the local ogi Salmonella, E.
coli and Shigella survived for 24 h or more but
showed a sharp decrease in numbers, while V.
cholerae survived for 12 h. DogiK is active
whether cooked or uncooked and exhibited
inhibition of pathogens at neutral pH. It gives
consistent quality. Preliminary investigation
indicates possession of a better shelf life.
Thus, DogiK may have a potential use in the
prevention and treatment of diarrhoea (Olukoya
et al. 1994).
10.6.8 Mahewu
Mahewu (amahewu) is an example of a non-
alcoholic sour beverage made from corn meal,
consumed in Africa and some Arabian Gulf
countries. It is known by various names, like
amahewu, amarehwu, emahewu, metogo, machleu
and maphulo; the most commonly used, however,
is mahewu. It is prepared from maize porridge,
which is mixed with water. Sorghum, millet malt
or wheat flour is then added and left to ferment
(Odunfa et al. 2001). Lactococcus lactis subsp.
Lactis, the natural flora of the malt, is the predomi-
nant micro-organism in the spontaneous fermenta-
tion carried out at ambient temperature (Steinkraus
1996; Gadaga et al. 1999).
Mahewu is known to offer some advantages
over ogi as the former consists of coarse maize
particles while ogi contains very fine pasty maize
particles. The initial wild fermentation by fungi,
in case of mahewu, is eliminated by boiling.
Furthermore, it is pre-cooked and requires only
mixing prior to consumption.
10.6.9 Pito
Pito is the traditional beverage, very popularly
consumed throughout Nigeria and Ghana owing
to its refreshing nature and low price. The
10 Maize Utilisation in Food Bioprocessing: An Overview
preparation of pito involves soaking cereal
grains, maize, sorghum or a combination of
both in water for 2 days. This follows malting
for 5 days in baskets lined with moistened banana
leaves. The malted grains are ground, mixed with
water and boiled. The resulting mash is allowed
to cool and later filtered through a fine mesh
basket. The filtrate thus obtained is allowed to
stand overnight, or until it assumes a slightly sour
flavour, following which it is boiled to a concen-
trate. A starter from the previous brew is added to
the cooled concentrate that is again allowed to
ferment overnight. Pito, the product thus
obtained, is a dark brown liquid which varies in
taste from sweet to bitter. It contains lactic acid,
sugars and amino acids and has an alcohol con-
tent of 3 %. Organisms responsible for souring
include Geotrichum candidum and Lactobacillus
sp. while Candida sp. are responsible for the
alcoholic fermentation.
10.6.10 Uji
It is primarily a sour maize gruel from East
Africa; however, alternatively some sorghum or
millet may also be added in 4:1 ratio (Mbugua
1984). The finely ground raw cereal is slurried
with water and allowed to ferment for 25 days at
25  C temperature. During fermentation, pH of
the slurry is reduced to 3.54.0 in 40 h. The
product is diluted to about 810 % solids and
boiled followed by further dilution and
sweetening with sugar before consumption. Uji
fermentation is characterised by the sequential
growth of the dominant micro-organisms,
initiated by the growth of coliforms and later
succeeded by the growth of lactic acid bacteria.
Early acid production at high concentrations by
the lactic acid bacteria rapidly restricts coliform
activity, thereby eliminating the problems of
off-flavours and flavour instability. Lactobacillus
plantarum is mainly responsible for souring of
uji, although early activity of heterofermentative
strains of L. fermentum, L. cellolbiosus and
L. buchneri during the fermentation is evident
(Onyango et al. 2003, 2004).
131
10.7 Limitations Associated with
the Use of Fermented Maize
Products
In most parts of Nigeria and Africa, children are
fed with mashed adult food or gelatinised cereal
flour slurries to complement breast milk from 4 to
6 months of age. These slurries absorb a large
quantity of water and swell up greatly when
mixed either with cold or hot water. The foods
are therefore bulky due to the high viscosity. Due
to the fact that high bulk reduces food intake by a
child often resulting in malnutrition, therefore,
development of nutritionally balanced calorie-
dense, low dietary bulk and easily digestible
weaning foods are being sought for. Moreover,
soaking of maize in alkali water (made with
ashes and calcium oxide) liberates the B-vitamin
niacin, the lack of which was the underlying cause
of the condition known as pellagra. Once alkali
processing and dietary variety were understood
and applied, pellagra disappeared in the developed
world. Besides the lack of niacin, pellagra was also
characterised by protein deficiency, a result of the
inherent lack of two key amino acids in premodern
maize, lysine and tryptophan. A range of indige-
nous fermented foods is prepared from cereals in
combination with legumes, thus improving the
overall protein quality of the fermented product.
Cereals are deficient in lysine but rich in cysteine
and methionine. Legumes, on the other hand, are
rich in lysine but deficient in sulphur containing
amino acids. Thus by combining cereal with
legumes, the overall protein quality is improved
(Campbell-Platt 1994). Consequently, number of
leguminous seeds including soya beans and okra
seeds are used to fortify and improve their protein,
iron, calcium and fibre content (Osungbaro 2009;
Anigo et al. 2009) to eliminate incidences of
anaemia and stunted growth, often associated
with malnutrition (Muhimbula and Issa-Zacharia
2010). Further, maize contains lipid transfer pro-
tein, an indigestible protein that survives cooking
and has been linked to a rare and understudied
allergy to maize in humans.
Above all, poor handling, inappropriate safety
measures and ill manufacturing practices will
132
ensure contaminated product that makes it source
of infections such as cholera, typhoid fever, hep-
atitis and gastroenteritis, in infants and adults.
For example, microbial assessment at critical
parts of ogi fermentation revealed contamination
of fungi like Aspergillus flavour, Aspergillus niger,
Penicillium oxalicum, Fusarium oxysporium and
Rhizopus stolonifer; yeast like Candida albicans
and Saccharomyces cerevisiae; enteric bacteria
like Escherichia coli and Klebsiella aerogenes;
and non-enteric bacteria like Lactobacillus
plantarum, Pseudomonas aeruginosa and Staphy-
lococcus aureus (Oyelana and Coker 2012). The
inclusion of fumonisin and aflatoxin in maize has
been linked to certain fungal species such as
Aspergillus, Penicillium, Rhizopus and Fusarium
(Omemu et al. 2005). The carcinogenic effect and
other health implications of these toxins were
extensively discussed by Shephard et al. (2002),
Barug et al. (2004) and Gadaga et al. (2004).
10.8 Conclusion
Fermented foods were discovered before mankind
had any knowledge of micro-organisms, but these
were simple observations that certain way of stor-
ing food effected desirable changes in its char-
acteristics such as shelf life. These characteristics,
in the industrial world, became less important
with the advent of alternative preservation
methods such as canning, chilling and freezing.
Modern technology, however, in no way has
diminished the sensory appeal, contributed by
peculiar flavour and aroma, of fermented
products. Fermented foods are considered benefi-
cial over non-fermented products as microbial
action not only improves shelf life by producing
acids, alcohols and bacteriocins but also provides
better digestibility, nutritional supplementation,
retention of micro- and macronutrients, improved
aroma and flavour characteristics to foods.
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 Maize Malting: Retrospect
and Prospect 11
D.P. Chaudhary, D. Kumar, R.P.S. Verma, Sapna Langyan,
and Seema Sangwan

Abstract
Malting is the process of converting cereal into malt. Barley is the most
preferred malted grain because of its high enzyme content needed for the
conversion of grain starch to malt. Wheat, rye, oats and rice are the other
cereal grains used for this purpose. Maize kernels are rich in starch
(70 %). The abundance of starch in maize stimulates researchers to
evolve improved technological interventions for the better conversion of
maize to malt. Inadequate diastatic power and the immature breakage of
plumule before the complete endosperm modification of maize kernel are
the major hurdles in selecting maize for malting purpose. Maize is used as
an adjunct in the production of beer. Breeding maize for malting and the
evolution of improved technology for proper endosperm modification are
needed for the efficient conversion of maize to malt.

11.1 Introduction amongst cereals as against the required growth

Maize is an important crop in the Indian agri-
cultural system that provides food, feed and
fodder and serves as a source of basic raw
material for a number of industrial products
including starch, oil, syrups, alcoholic
beverages and biofuel. In the last 5-year plan,
maize achieved the highest growth rate (6.7 %)
D.P. Chaudhary (*)  S. Langyan
Directorate of Maize Research, New Delhi, India
e-mail: chaudharydp@gmail.com
D. Kumar  R.P.S. Verma
Directorate of Wheat Research, Karnal, India
S. Sangwan
Division of Microbiology, CCS HAU, Hisar, India
rate of 4.7 % set by the planning commission of
India. The production as well as productivity of
maize is increasing continuously for the last
few years. As a result, India moved from
being importer to exporter of maize. Consider-
ing the changing climatic scenario and the
impact of single-cross maize hybrids, it is
estimated that production and productivity of
maize will further increase in the near future. In
the present utilization pattern, a major part of
maize produced is used as animal feed followed
by food. A significant amount of maize pro-
duced is used as an industrial raw material. At
present, around 20 % of maize produced is
utilised in the starch industry for the production
of starch along with maize oil, bran and gluten
meal as by-products. As a result of increasing

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 135
DOI 10.1007/978-81-322-1623-0_11, # Springer India 2014
136 D.P. Chaudhary et al.

maize production, its industrial utilisation is Table 11.1 Chemical composition of the main parts of
also increasing, resulting in further expansion maize kernels (%)
of this segment. Since maize is one of the most Chemical
important multipliers of starch in this universe, component Pericarp Endosperm Germ
its use as a raw material in the malting industry Protein 3.7 8.0 18 4
could effectively be explored. To date, barley Ether extract 1.0 0.8 33.2
Crude fibre 86.7 2.7 8.8
is primarily being utilized for malting purpose.
Ash 0.8 0.3 10.5
The abundance of starch in maize, however,
Starch 7.3 87.6 8.3
prods the researchers to explore improved tech-
Sugar 0.34 0.62 10.8
nological interventions as well as instrumenta-
Source: Watson (1987)
tion for the efficient conversion of maize into
malt. In this chapter the nutritional quality of
maize is being discussed followed by its Table 11.2 Gross chemical composition of the different
types of maize (%)
strengths as well as weaknesses with respect
to its conversion to malt for brewing purpose. Maize type Protein Sugar Oil Starch
Normal (HM 4) 11.8 3.3 3.5 70.3
QPM (HQPM 4) 10.8 3.9 3.3 67.7
11.2 Structure of the Maize Kernel Sweet (WOSC) 13.6 13.4 12.1 50.0
Pop 13.7 2.5 5.7 66.0
High oil (HKI-162-2) 11.0 3.5 6.2 69.9
Maize kernels vary in size and shape in different
kinds and varieties. They may be only an eighth of Source: 53rd Annual Progress Report, DMR, (2010);
Cortez and Wild-Altamirano (1972)
an inch long and near round in popcorn to a half-
inch long and a flattened-cylinder shape in some
other kinds. Maize kernels develop on the female
inflorescence called the ear. The ear is a long components are considered indigestible for
cylindrical structure that may hold from 200 to humans but help in the proper movement of
1,000 single kernels depending on the number of bowl. However, ruminants can efficiently digest
rows, diameter and length of the cob. The number the bran and, therefore, it is used in animal feed.
of rows may vary from 12 up to 22 depending upon Endosperm, the major component of maize ker-
the genotype, as well as quantum of nutrients nel, contains high levels of starch (8588 %)
applied. There are four major physical structures and protein (8 %). The germ is rich in fat,
of the kernel: the outermost called pericarp, hull or averaging about 30 %. It also contains relatively
bran; the germ or embryo; the endosperm; and the high levels of protein (18 %), sugar (11 %)
tip cap (dead tissue found where the kernel joins and some minerals.
the cob). The endosperm is the largest component The chemical composition, however, varies
comprising about 8085 % of the kernel weight, amongst different types of maize. A wide
whereas germ is about 10 % and the pericarp variability is observed in each major nutrient
around 5 % of the kernel weight. component. Table 11.2 summarises data on vari-
ous types of maize. The variability observed is
both genetic and environmental. It may influence
11.3 Chemical Composition the weight distribution and individual chemical
composition of the endosperm, germ and hull of
The different components of maize kernel differ the kernels.
significantly in terms of chemical composition Starch is the major carbohydrate of the maize
(Table 11.1). The pericarp is characterised by a kernel, averaging around 6774 %. Other
higher concentration of crude fibre (87 %), carbohydrates are simple sugars, mainly sucrose
consisting mainly of hemicellulose (67 %), cel- and very small amounts of glucose and fructose.
lulose (23 %) and very small quantities of lignin Starch is an odourless, white powder with a bland
(0.1 %) (Burge and Duensing 1989). These taste, which is extensively used as an industrial
11 Maize Malting: Retrospect and Prospect
material for the synthesis of a number of
products. Maize is one of the most important
multipliers of commercially produced starch.
Chemically, starch is a polysaccharide consisting
of a large number of glucose units joined by
glycosidic bonds. It consists of two types of
molecules: the amylose and the amylopectin.
Amylose is a linear polymer in which the -D
glucose units are linked through -1,4 glycosidic
bonds. Amylopectin is a branched structure in
which apart from -1,4; -1,6 glycosidic bonds
are also present. The composition of maize starch
is genetically controlled. In normal maize, amy-
lose constitutes up to 2530 % of the starch and
amylopectin up to 7075 %. Maize that contains
100 % amylopectin is called waxy maize. An
endosperm mutant called amylose-extender (ae)
has an increased amylose proportion of the starch
induced up to 50 % and higher. Other genes,
alone or in combination, may also modify the
amylose-to-amylopectin ratio in maize starch
(Boyer and Shannon 1987). The waxy as well
as ae lines have different applications in the
starch industry. Waxy maize is mainly utilised
in the food industry primarily due to its viscous
and gel-forming properties. High amylose maize,
on the other hand, is hard due to its compact
chemical structure and is used for making starch
threads, etc. Starch is extensively used in the
food, pharmaceutical, textile, paper, adhesive
and a host of other industries. It can also be
converted into a range of starch derivatives
such as liquid glucose, dextrose monohydrate,
dextrose anhydrous, maltodextrin and sorbitol.
Protein is the second largest and an important
component of maize kernel. In normal maize, the
protein content varies from 7 % to 13 % of the
kernel weight. Most of the protein is located in the
endosperm. It is made up of at least five different
fractions (Landry and Moureaux 1970, 1982).
Albumins, globulins and non-protein nitrogen
amount to 18 %, whereas prolamine fraction
contributes 52 % of the nitrogen in the kernel.
Prolamine 1 or zein 1 is found in the largest
concentration (42 %), whereas prolamine 2 or
zein 2 is found to be around 10 %. Glutelin
fraction 2 amounts to about 8 %, while glutelin
3 amounts to about 17 % for a total glutelin
content of 25 % of the protein in the maize
137
kernel (usually a small amount, about 5 %, is
residual nitrogen). In the normal maize, zeins
predominate in the endosperm, which are primar-
ily responsible for the poor quality of protein of
normal maize. To the contrary, the quality pro-
tein maize is characterised by higher content of
non-zeins, which are better balanced in terms of
protein quality.
Although a by-product, corn oil occupies a
significant position in human nutrition. The oil
comes mainly from the germ. Its concentrations
are genetically controlled, with values ranging
from 2 % to 20 %. Maize oil has low level of
saturated fatty acids. On the other hand, it
contains relatively high levels of polyunsaturated
fatty acids, mainly linoleic acid (24 %). Very
small amounts of linolenic and arachidonic
acids have been reported. Furthermore, maize
oil is rich in vitamin E, which makes it relatively
stable. Maize oil is highly regarded for human
consumption because of its fatty acid profile,
mainly oleic and linoleic acids.
Dietary fibre consists of indigestible portions
of carbohydrates. It comes mainly from the
pericarp and the tip cap, although small
quantities are also contributed by endosperm
cell walls and germ cell walls. Maize bran is
composed of 75 % hemicellulose, 25 % cellulose
and 0.1 % lignin on a dry-weight basis. Small
amounts of sugars (26 %) are also found in
mature maize kernel with sucrose, the major
component, found mostly in the germ. Higher
levels of monosaccharides, disaccharides and
trisaccharides are present in maturing kernels.
Apart from this, minerals also contribute
between 1 % and 2 % of the kernel weight.
Germ provides about 78 % of the whole kernel
minerals. The most abundant mineral is phospho-
rus, found as phytate of potassium and magne-
sium. All the phosphorus is found in embryo.
Maize is low in calcium and trace minerals.
11.4 Malting
In the simplest term, malting is the controlled
germination and kilning of grain. The grains are
made to germinate by soaking in water and are
then halted from germinating further by drying
138
with hot air. Malting grains develops the enzymes
(diastatic enzymes) required to modify the grain
starch into sugars, including monosaccharides,
such as glucose or fructose, and disaccharides,
such as sucrose or maltose. It also develops other
enzymes, such as proteases, which break down
the proteins in the grain into molecules that can
be used by yeast. The malting of cereal grains
involves controlled germination, and finally
biological activities are arrested through gradual
heating. Though, the most preferred grain for
malting is barley, but other grains like wheat,
maize and sorghum are also used for making malt.
11.5 Malting Process
Malting is a three-step process consisting of
steeping, germination and finally kilning. A
brief detail of all the three steps is outlined in
the following:
11.5.1 Steeping
The basic aim of this process is to increase the
moisture content of the grain while maintaining
seed viability. This is required for the germina-
tion of the grain and for uniform diffusion of
enzymes throughout the endosperm. On an aver-
age, the moisture content is raised to more than
40 %. This operation is done in conical vessels or
flat-bottomed tanks. Conical vessels have limited
capacity of around 50 t, while flat-bottomed
tanks have capacity of around 300 t. The grains
are initially dipped in water, and a temperature of
1418  C is maintained for about 68 h. During
this step, the dirt, floating kernels and some
undesirable compounds get removed. Since the
metabolic activity in the grain gets started during
this period, the dissolved oxygen in the steep
water gets depleted. To supply the oxygen and
to remove the excess carbon dioxide, air is bub-
bled through water, and after the immersion
cycle, water is drained and fresh air is circulated,
and this phase is known as the air rest stage. The
air rest stage is followed by again steeping or
D.P. Chaudhary et al.
immersion stage and then again air rest stage.
The duration of each stage and total steeping
depends upon the kind of grain and variety of
grain being used, as at the end of the steeping
period, the grain is usually in chitted state.
11.5.2 Germination
This process is done to achieve modification of
the endosperm reserves, especially starch. Dur-
ing this process, hydrolytic enzymes, that is,
amylolytic and proteolytic ones, become active
and break down complex molecules into simple
ones. The cell walls of endosperm cells also get
disintegrated during this process. The commercial
germination unit consists of a large compartment
to accommodate 50300 t of grains, with a provi-
sion to provide cool and humidified air and slow
turning of grains. Turning of grains is essential to
separate the growing roots so that the flow of air
through the bed is uniform. Temperature is kept
between 16 and 18  C to minimise the respiratory
losses, which will ultimately lead to decreased
malt yield. Again germination period varies from
3 to 7 days depending upon the kind of grain and
variety being used.
11.5.3 Kilning
The major objective of kilning is to dry the
germinated grains to the moisture content of
45 % and to achieve development of malt
colours and flavours. The kiln has provision of
gradual temperature increase and air circulation,
and energy is conserved to the maximum extent
possible. The kilning is usually done for 1640 h
with gradual increase in the temperature. The
temperature is raised from 45  C to 7080  C
gradually to preserve the enzyme activities. Dur-
ing the kilning process, the temperature of grains
is usually lower than the heating air temperature
because of moisture loss from the grains. After
the kilning process, the grains are cooled and
rootles removed. The malt is cleaned and packed
in airtight bags for shipment.
11 Maize Malting: Retrospect and Prospect
11.6 Maize Malting: Problems
and Future Prospective
The malting potential of different cereals has
been studied over time (Malleshi and Desikachar
1989). Barley is the most preferred grain for
brewing all over the world. It is the fourth largest
cultivated cereal crop in the world. There are
many advantages of barley over other cereal
grains for malting process. It has a short germi-
nation period suited for malting process. It has
sufficiently high starch content and low protein
which reduces haze in beer. However, the most
important characteristic that makes it suitable for
malting is perhaps its high diastatic power (high
level of diastatic enzymes) needed for conversion
of starch (Kneen 1944; Engel 1947; Norris
and Lewis 1965; Palmer 1989; Muoria 1998).
Secondly, barley grain is covered by husk
which has advantage in malting since during
germination, the endosperm modification is
determined by the length of the acrospire. The
growth of the acrospire is used as a rough guide
to the progress of malting. Usually, the lengths of
the acrospires of a number of grains are noted as
fractions of the grain lengths. Thus, in traditional
practice the green malt is kilned and growth
terminated when most acrospires are 0.751 of
the grains length. In barley, the acrospires
develop beneath the husk, whereas in corn the
plumule sprouts upward making it vulnerable to
break immaturely before the completion of the
endosperm modification process. The steeping
and germination is usually carried out in big
tanks where a grain bed is formed for this pro-
cess. The grain bed is continuously agitated to
avoid deoxygenation. During the process, the
breakage of plumule is the most common prob-
lem with cereals other than barley.
The second important disadvantage with corn
is its diastatic power. In a preliminary study
carried out at the Directorate of Wheat Research,
Karnal, it was observed that corn produces
around 5060 % of malt after steeping and ger-
mination period spanning 67 days (unpublished
data). The maize kernel consists of hard endo-
sperm, which is surrounded by a thin and
139
uniform layer of pericarp. This increases the
steeping period. Usually, it takes around 34
days to gain maximum moisture content, and
the total period for germination is around a
week. To date, corn is being used as adjunct
along with barley in the malting process (Iwouno
2012). In order for enzymatic conversion to
occur, grain starches must be gelatinised, which
means their structure must be broken down by
heat and water. Each grain has a different tem-
perature at which gelatinisation occurs. Corn has
a higher gelatinisation temperature than barley,
which needs special processes to make the starch
available for conversion. Maize generally has little
or no diastatic power and so must be used with
base malts for starch conversion (Takaku 1988).
Malting of maize for use as a major source of
hydrolytic enzymes required for brewing purposes
has received less attention (Eneje et al. 2004).
Earlier works have shown that cereal grains
could differ significantly in their modes of
amylases development (Kneen 1944; Lauriere
1992; Ziegler 1999). The low activity of amylases
in most cereals other than barley malt has been
defined as one of the most serious obstacles to
their use as replacement for barley malt (Takaku
1988; Okungbowa et al. 2002). However, the
gluten-free characteristics of maize make it a good
substitute for barley malt especially for people
suffering from celiac disease (Sweeney 2004).
However, considering the magnitude of starch
available from maize, efforts are continuously
being made to explore the experimental variables
for better maize malting. In an earlier study, 27
yellow maize varieties were malted using varying
steeping time (30 h, 36 h, 42 h), germination time
(3 days, 4 days, 5 days) and kilning temperature
(50  C, 55  C, 60  C). Results revealed that 42 h
of steeping, 5 days of germination and 50  C
kilning temperature give the best maize malt
(Wouno and Ojukwu 2012). However, maize
malt showed low diastatic power when compared
with barley malt as reported by Okafor and
Aniche (1980).
The use of maize as malting grain attracts little
attention in India. However, with the increasing
production and productivity of maize, a lot of
opportunities are available whereby we can
140
explore the possibility of a good quality beer
produced from corn. Since maize possesses a suf-
ficiently high quantity of starch, efforts could be
diverted towards exploring new ideas and scien-
tific know-how to use maize as an efficient grain
towards better beer production.
References
Annual Progress Report (2010). Directorate of Maize
Research. Indian Council of Agricultural Research,
New Delhi.
Boyer CD, Shannon JC (1987) Carbohydrates of the ker-
nel. In: Watson SA, Ramstad PE (eds) Corn: chemis-
try and technology. American Association of Cereal
Chemists, St Paul, pp 253272
Burge RM, Duensing WJ (1989) Processing and dietary
fiber ingredient applications of com bran. Cereal
Foods World 34:535538
Cortez A, Wild-Altamirano C (1972) Contributions to the
lime-treated corn flour technology. In: Bressani R,
Braham JE, Behar M (eds) Nutritional improvement
of maize, vol L4, INCAP Publication. INCAP,
Guatemala, pp 99106
Eneje LO, Ogu EO, Aloh CU, Agu RC, Palmer GH (2004)
Effects of steeping and germination on malting perfor-
mance of Nigerian white and yellow maize varieties.
Process Biochem 39:10131016
Engel C (1947) The distribution of enzymes in resting
cereals. Biochem Biophys Acta 1:4249
Iwouno JO, Ojukwu M (2012) Effect of experimental
variables on the malting qualities of Nigerial yellow
maize (Zea mays) farz 27 variety. Afr J Food Sci
3:252259
Kneen E (1944) A comparative study of the development
of amylases of germinating cereals. Cereal Chem
21:304314
D.P. Chaudhary et al.
Landry J, Moureaux T (1970) Heterogeneite des glutelines
du grain de mas: extraction selective et composition en
acides amines des trots fractions isolees. Bull Soc Chim
Biol 52:10211037
Landry J, Moureaux T (1982) Distribution of amino acid
composition of protein fraction in opaque-2 maize.
Phytochemistry 21:18621869
Lauriere C, Doyen C, Tevenenot C, Daussant J (1992) A
study of the Maize Amylase. Plant physiol 100:877
Malleshi BG, Desikachar HSR (1989) Studies on compar-
ative malting characteristics of some tropical cereals
and millets. J Inst Brew 92:174176
Muoria JK, Bechtel PJ, Linden JC (1998) Diastatic
power and -Amylase activity in millet sorghum and
barley grains and malts. J Am Soc Brew Chem
56:131135
Norris K, Lewis M (1965). Technical Quarterly Associa-
tion of the Americans Master Brewers. 22: 154
Okafor N, Aniche GN (1980) Brewing lager beer from
Nigerian sorghum. Brewing Distilling Int 10:3233
Okungbowa J, Obeta N, Ezeogu L (2002) Sorghum beta
amylase production. J Inst Brewing 108:362370
Palmer GH (1989) Cereal science and technology.
Aberdeen University Press, Aberdeen, pp 147148
Sweeney (2004). Gluten free brewing. http://www.living.
fortunecity.Com/boozersbrewers
Takaku H (1988) Handbook of amylase and related
enzyme: their sources isolation methods properties
and application. (Amylase research society of Japan).
Pergamon Press, New York, pp 215217
Watson SA (1987) Structure and composition. In: Watson
SA, Ramstad PE (eds) Corn: chemistry and technology.
American Association of Cereal Chemists, St Paul,
pp 5382
Wouno JO, Ojukwu M (2012) Effects of experimental
variables on the malting quality of Nigerian yellow
maize (zea Mays), farz 27 variety. Afr J Food Sci
Technol 3:252259
Ziegler P (1999) Cereal beta amylase. J Cereal Sci
29:195204
 Value Addition in Maize
12
V.K. Yadav and P. Supriya

Abstract
Globally, maize is the most important coarse grain cereal and well known as
poor mans nutricereal due to presence of high content of carbohydrates,
fats, proteins, and some of the important vitamins and minerals. On the basis
of its unique characteristics and nutritional composition specialty, corn is
classified into quality protein maize, baby corn, sweet corn, pop corn, green
eared corn, high oil corn, etc. Specialty corn has diverse end uses and a
number of value-added products can be prepared from it. Quality protein
maize differs from normal maize as it contains added amount of essential
amino acids such as tryptophan and lysine. Baby corn is the ear, harvested
young when the silks have just emerged and no fertilisation has taken place.
In sweet corn, the taste of kernels is much sweeter than normal corn. In
popcorn when kernels are heated, they explode and produce large puffed
flakes (popping). The value-added products prepared from specialty corns
are traditional foods, infant foods, health foods, snacks and savory, baked
products, etc. Each specialty corn has different recipes. Apart from these
products, maize is used to prepare industrial products such as starch, specialty
chemicals, ethanol, refined corn oil, sorbitol, cake mixes, candies, carbonated
beverages, and cosmetics.

12.1 Introduction the important vitamins and minerals, maize has

Maize [Zea mays L.] is a major cereal crop
for human nutrition. With its high content
of carbohydrates, fats, proteins, and some of
V.K. Yadav (*)
P. Supriya
Directorate of Maize Research, Pusa Campus,
New Delhi 110012, India
e-mail: vkyadavdmr@rediffmail.com
acquired a well-deserved reputation as poor
mans nutricereal. Several million people, par-
ticularly in the developing countries, derive their
protein and calorie requirements from maize.
The incomparability of corn is due to its wide
range of uses and highly useful products into
which it can easily be transformed. Maize crop
has a special place in Indian agriculture and is
a staple food of people of low socio-economic
groups.

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 141
DOI 10.1007/978-81-322-1623-0_12, # Springer India 2014
142 V.K. Yadav and P. Supriya

Table 12.1 Nutritional composition of quality protein maize and normal maize (dry matter basis)
Particulars HQPM-1 Shaktiman-1 HM-4 HM-5
Moisture (g/100 g) 6.90 7.34 7.37 7.23
Crude protein (g/100 g) 11.48 11.65 10.04 10.15
Crude fat (g/100 g) 4.40 4.52 4.43 4.29
Crude fibre (g/100 g) 2.41 2.34 2.40 2.65
Ash (g/100 g) 1.46 1.55 1.34 1.58
Total carbohydrates units 80.28 79.94 81.97 81.33
Energy (Kcal/100 g) 465.2 407.04 375.67
Total soluble sugars (g/100 g) 2.34 2.81 0.14 2.58
Calcium (mg/100 g) 15.39 17.78 17.76 17.53
Phosphorous (mg/100 g) 200.63 200.78 197.89 197.63
Iron (mg/100 g) 2.74 2.78 2.73 2.50
Source: Kawatra and Sehgal (2007)

Corn is largely classified into six types based
upon the quality, quantity, and composition of
endosperm. These types are known as dent, flint,
flour, sweet, pop and pod. Each has different
properties with various end uses. In India, much
work has been done in the area of specialty corns
such as sweet corn, popcorn, baby corn, quality
protein maize, green-eared corn, high-oil corn,
waxy corn and fodder maize.
12.2 Quality Protein Maize and
Its Value-Added Products
In spite of diverse uses, maize has many
drawbacks, especially imbalance in amino acids
composition. To overcome these problems,
scientists have discovered the maize mutant
opaque-2, which is superior in its nutrient com-
position than common maize and is named as
quality protein maize (QPM).
QPM itself is value-added maize as it contains
increased amount of essential amino acids such
as tryptophan and lysine. The discovery of QPM
was initiated in the early 1960s, when scientists
at Purdue University discovered a peculiar gene
that significantly increased the level of two
essential amino acids, lysine and tryptophan, in
the maize grain. QPM looks and tastes like
normal maize and yields as much or more, but
it contains nearly twice the quantity of the
essential amino acids, lysine and tryptophan,
which are essential building blocks of proteins
in humans and monogastric animals like poultry
and pigs (CIMMYT 2000).
The QPM has better protein quality than com-
mon maize because it contains double the
amount of lysine and tryptophan and no change
in other amino acids except low level of leucine.
Decrease in leucine is considered particularly
desirable as it makes the leucineisoleucine
ratio more balanced, which in turn helps to liber-
ate more tryptophan for niacin biosynthesis and
thus helps to combat pellagra (Prasanna et al.
2001). Other nutritional benefits of QPM include
higher niacin availability due to higher trypto-
phan and lower leucine content, higher calcium
and carbohydrate and carotene utilisation.
12.2.1 Nutrient Composition
of Maize/QPM
A number of QPM varieties are being
released with wide variability in nutrient com-
position. Two varieties of QPM (HQPM-1 and
Shaktiman-1) and two varieties of normal maize
(HM-4 and HM-5) have been analysed at
CCSHAU Hisar for their nutrient profile, which
is presented in Table 12.1. Protein, fat and ash
content ranged from 10 to 11 %, 4.25 to 4.53 %,
and 1.34 to 1.58 %, respectively.
12 Value Addition in Maize 143

Fig. 12.1 Value-added products from blends of maize and legume

12.2.2 Value-Added Products Snacks and QPM biscuit salted, QPM biscuit
of Quality Protein Maize savoury item sweet, choco maize biscuit, honey
maize chikki, maize matthi, namak
para, sev, shakarpara, QPM burfi,
In India, maize is generally consumed in the QPM halwa, suji upma, suji kheer,
form of chapati, popcorn, roasted fresh cob, etc. sevian (sweet), sevian (upma),
A wide variety of products (Fig. 12.1) have been QPM chatni powder-I, QPM
developed in the Department of Food and Nutri- chatni powder-II and QPM chatni
powder-III
tion, CCSHAU, Hisar, RAU, Pusa (Bihar) and
Specialty foods High-quality protein mix,
UAS, Mandya (Karnataka) which can meet the low-quality protein mix, quality
nutritional need of the vulnerable section. To protein mix for elderly, QPM honey
further improve the nutritive value of the QPM, liquid, and honey maize water
legumes can be supplemented to prepare blends (Singh 2006)
to improve the protein quality of the products.
Many products have been developed by using
flour blends prepared with addition of soya Quality Protein Maize was blended with soya
bean and green gram: bean/green gram in the ratio 70:30 and the prod-
uct developed includes the following:
Traditional Ladoo, halwa, kheer, chapati, sev,
products mathi, pakora and cheela
Baked products Bread, nan khatai and cake
Traditional products Cake, biscuit
Extruded Vermicelli and pasta
Baked products Halwa, upma, dalia,
products
cheela, namak para,
Convenience Instant idli mix, instant dhokla mix, sattu, khichri, burfi
foods and porridge mix; sprouted products
Dehydrated products Vadi and fryums
sprouted chat, QPM vada, QPM
sevian and QPM flour
Infant food Infant food-I, infant food-II, infant
food (flavoured), infant food
(enriched with vitamin A) and infant All the products have been found acceptable
food (flavoured and enriched with to people in the acceptability trials. Value-added
vitamin A) products were nutritious, rich in protein, energy,
Health food QPM mix-I, QPM mix-II, QPM fat, etc. Products made with addition of legumes
ladoo, honey maize chocolate,
maize coconut chocolate, maize
have better protein in terms of both quantity and
coconut toffee, maize groundnut quality. The products can be stored well up to
toffee, choco maize bar and honey period depending on nature of the product
maize water (Kawatra and Sehgal 2007).
144 V.K. Yadav and P. Supriya

Table 12.2 Nutritional composition of baby corn (fresh

12.3 Baby Corn and Its matter basis)
Value-Added Products Parameters Content (g/100 g)
Proximate composition
Baby corn, as the name suggests, is the young corn Moisture 90.03  0.16
cob at its earliest stage of development. Baby corn is Crude protein 17.96  0.40
the ear, harvested young, when the silks have either Crude fat 2.13  0.13
not emerged or just emerged and no fertilization has Total ash 5.30  0.08
taken place. Baby corn is a very delicious and Crude fibre 5.89  0.34
Available carbohydrates
nutritious vegetable and has been considered to be
Total soluble sugars 23.43  0.23
a high-value agriculture produce in national and
Reducing sugars 1.96  0.03
international markets. The story of its production
Non-reducing sugars 21.47  0.13
technique is approximately three decades old when
Starch 15.60  0.02
in 1972, the government of Thailands agricultural -carotene (mg/100 g) 670
research policy focused on baby corn research and Ascorbic acid (mg/100 g) 5.43
development in order to improve the deteriorating Phosphorous (mg/100 g) 898.62
economy of the country. As a result, in a record Source: Kawatra and Sehgal (2007)
time, this crop became popular in the whole of
Thailand and the farmers collected valuable foreign 12.3.1.1 Canning
exchange through its production. In India, baby Canning is a commonly used method for
corn industry is still at the juvenile stage. At present, processing baby corn. Baby corn can be canned
baby corn production is confined to peri-urban areas in brine, stored for months together, and
as its consumption is mostly confined to big hotels transported to far-off places. Farmers can be
and restaurants. The nutritive value of baby corn is trained to establish the canning plants or made
comparable to several high-priced vegetables like aware regarding various processing units avail-
cauliflower, cabbage, French bean, spinach, lady able in the nearby areas, so that they can get good
finger, brinjal, tomato and radish. Besides being price for their crops. Canning is a simple process
nutritious, it has several other advantages too. It is following a sequence of steps:
free from the harmful effects of pesticides as baby
corn is covered tightly with husk and due to early
Peeled baby corn ! cleaning ! boiling
harvesting; it is devoid of harmful pests and
diseases. In addition, baby corn has a great potential ! soaking ! grading
for providing higher economic benefits to farmers ! canning ! brining
as it is possible to have three or more crops from the ! exhausting ! lid covering
same piece of land as this crop matures in less than ! cooling ! quality inspection
60 days during kharif and farmers also get good
price for it. Baby corn is highly nutritive as it
contains sufficient quantities of carbohydrates,
12.3.1.2 Dehydration
proteins, minerals, and vitamins, particularly beta
Dehydration can be used to increase the shelf life
carotene (Table 12.2).
of baby corn for a long period. Baby corn can be
cut into 0.5 cm round pieces and dried (in air
oven, vacuum oven, or can be solar dried). Dried
12.3.1 Processing of Baby Corn baby corn packed in polythene pack/vacuum
pack/tetrapack can be stored well for a longer
Baby corn can be processed to improve its shelf period. Shelf life of dry baby corn stored for 3
life. Main processing methods which can be used months has been studied on trial basis. Non-
to improve shelf life are as follows. significant changes in moisture, crude protein,
12 Value Addition in Maize
Fig. 12.2 Baby corn products
crude fat, total ash, crude fibre, minerals, protein
content and starch digestibility have been
observed after 3 months. Dehydration results
in significant losses of ascorbic acid, which
further reduces during storage of dehydrated
baby corn. Similarly, -carotene content is also
lost during dehydration and storage of baby
corn. Dehydrated baby corn can be rehydrated
by soaking in water and can be used in prepara-
tion of food products. Products developed using
dried baby corn have been found acceptable
organoleptically, like those prepared from fresh
baby corn.
12.3.1.3 Freezing
Baby corn can be frozen and stored for long period
like other frozen vegetables, maintaining cold
chain. Storage of frozen baby corn for 3 months
produced non-significant changes in proximate
principles. Storage of frozen baby corn in cold
storage produced significant reduction of 6.95 %,
9.02 % and 11.06 % in ascorbic acid content on
30th, 60th and 90th day of storage, respectively. -
carotene content has been found to be reduced by
1011 % on 3 months of frozen storage. Frozen
baby corn can be used effectively for preparation
of food products. Preparations like soups and
vegetables prepared from frozen baby corn are as
acceptable as from fresh baby corn. Frozen baby
corn can be directly used in various preparations.
145
12.3.2 Value-Added Products
of Baby Corn
Baby corn can be eaten raw. It can be included in
diets in a number of ways as salads, Chinese and
Italian preparations. A wide range of food
products have been developed at CCS HAU,
Hisar, which are not only acceptable but can be
easily prepared at home (Fig. 12.2):
Traditional Pakoda, cutlet, chat, salad, dry vegetable,
products kofta, mixed vegetable and raita
Sweet Halwa, kheer and burfi
products
Preserved Jam, chutney, pickle, candy and
products murraba
Chinese Soup, Manchurian, baby corn chilli,
products chow mein, and sweet and sour
vegetable
12.4 Sweet Corn and Its
Value-Added Products
Sweet corn (Zea mays L. saccharata) is one of
the most popular vegetables in countries like the
USA and Canada. It is becoming increasingly
popular in India and other Asian countries. It is
consumed in immature stage of the cob. Sweet
corn varies from normal corn essentially due to
gene(s) that affects starch synthesis in the seed
endosperm wherein one or more simple recessive
146 V.K. Yadav and P. Supriya

alleles alter the carbohydrate content of the endo-
sperm and elevate the level of water soluble
polysaccharides (sugars) and decrease starch.
Thus, the kernels of sweet corn taste much
sweeter than normal corn, especially at 1821
days after pollination. The total sugar content in
sweet corn ranges from 25 % to 30 %. Sweet corn
matures early and can be harvested in 7580 days
after planting. Thus, it can fit easily in multiple or
intercropping systems. Due to the absence of
starch in the kernels (seeds), the dried seed is
shriveled in appearance.
Modern sweet corn cultivars arose in the nine-
teenth century when a single gene (su) mutated in
field corn. Plants descending from this mutant had
kernels with a sugary coating rather than a peri-
carp (seed coat) than normal corn, making it
tender. In earlier history of sweet corn, lines
with only the sugary (su1) allele on chromosome
4 used to be referred to as sweet corn. Currently,
at least eight genes that affect carbohydrate syn-
thesis in the endosperm are being used either
singly or in combination to breed for sweet corn
varieties. Depending on the gene combination
used, endosperm types differ in eating quality,
shelf life of both fresh and processed sweet
corns, and their field emergence. Thus, they
often require specialised seed production, plant-
ing techniques, and either distance or time isola-
tion to prevent cross-pollination. These types are
referred to as super sweets, although technically
this term only describes cultivars with the
shrunken 2 (sh2) characteristic.
12.4.1 Value-Added Products
of Sweet Corn
12.4.2 Future Strategies for Sweet
Corn in India
In spite of availability of promising varieties in
India and potential of the technology, sweet corn
has yet to assume popularity in the country. A
major portion of the sweet corn is being utilised
in hotels and big restaurants. The main reason
behind this is non-existence of appropriate stor-
age and marketing facilities in India. Thus, there
is an urgent need to develop entrepreneurship to
harness the advantages associated with sweet
corn. Since sweet corn needs to be utilised imme-
diately after harvest, its cultivation should be
encouraged in peri-urban regions of the country.
Market network must be developed to ensure
household availability of sweet corn. This can
only be attained with proper market leadership
and policy support, which is presently lacking in
India (Rakshit et al. 2003).
12.5 Popcorn and Its Value-Added
Products
Popcorn (Zea mays L. indurata) is a popular
snack food throughout the world. Kernels of
popcorn range in colour from off-white gold to
red, black and many colours in between. When
kernels of popcorn are heated, they explode and
produce large puffed flakes (popping) (Fig. 12.3).
Once popped, popcorn has two basic shapes:
snow flake or butterfly, which pops big and
shapes like a cumulus cloud, and mushroom,

A number of sweet corn products have been devel-

oped for commercial use. Some of these include:
Thai basil and sweet corn
Salted green beans with shallots and sweet corn
Organic speed chefpizza with garlicky
greens and sweet corn
Sweet corn cake
High summer scallops with sweet corn and
couscous
Sweet corn and tomato salad Fig. 12.3 Popcorn
12 Value Addition in Maize
which pops into a round ball. An added feature of
popcorn is its light and crunchy texture.
Popcorn is used primarily for human con-
sumption as fresh or as the basis of its
confections. Besides their use as a popular
snack, ground popcorn as flour or grits can be
used in the preparation of many traditional
dishes. Isolated planting is not necessary in case
of popcorn, since there are no major xenia effects
on popping expansion and much popcorn are
cross-sterile with field corn. Although the
conditions for growing popcorn are the same as
for the dent corn, special harvesting, dry, and
storage practices are necessary to maintain
popping quality.
Popcorn plant type has some distinctive
characters compared to normal corn. The plant
type is lanky. The tassel is highly branched and
the branches are droopy. The ear placement is
higher up compared to normal corn. Often the
prolificacy is more in fertile soils with good
management. It is very common to find two
ears per plant. In many popcorn types, there is
tendency of tillering, and the brace roots are also
fewer compared to normal corn. In popcorn, sev-
eral grain colours are grown, viz., white, yellow,
and red. However, yellow types are more com-
mon. There are two main types of popcorn: rice
type and pearl type. The rice-type popcorn
kernels are common in white-grain types. They
are typically beaked, that is, long and pointed at
the tips. Pearl-type popcorn is more common than
the rice type. It has smooth and round kernels and
is common in yellow-grain type. Based on the
type of flakes produced on popping, popcorn is
further classified as butterfly type (flat flakes) and
mushroom types (compact globose flakes).
12.5.1 How Does Popcorn Pop?
The nutritional composition of popcorn kernel
consists of carbohydrate (principally starch), pro-
tein, fat, and water. Water is stored in a small
circle of soft starch in each kernel. As kernel is
heated, the water heats, builds up pressure, and
takes up any available room. The harder surface
surrounding the starch resists the vapour pressure
147
for as long as it can. When the outer surface gives
way, the water vapour further expands, causing
popcorn to explode. The soft starch pops out, the
kernel turns inside out, steam inside the kernel is
released, and the corn pops.
12.5.2 Nutritional Value of Popcorn
Popcorn is a good source of carbohydrate, energy
and fibre. For individuals of normal weight, car-
bohydrate is the best source of body fuel. Pop-
corn is a rich source of fibre, which is an
important component of human diet. There are
two types of fibre: (i) soluble fibre which plays a
role in regulating hunger, cholesterol and blood
sugar; and (ii) insoluble fibre, which is important
in gastrointestinal functioning. Popcorn has no
artificial colour or flavour additives and is sur-
prisingly low in calories. One cup of popcorn has
31 cal if eaten plain or seasoned with herbs;
133 cal if drizzled with a tablespoon of butter,
margarine, or oil; 34 cal if sprayed with butter-
flavoured oil; and 35 cal if sprinkled that butter
substitute. It is a wholesome, fun food that aids
digestion by providing necessary roughage.
Health and medical associations regard popcorn
as an excellent mealtime complement: sugar-
free, fat-free and low in calories. The National
Cancer Institute (NCI), USA, suggests that
increased fibre intake is associated with reduced
incidence of cancer. The American Dental Associ-
ation includes popcorn in its list of recommended
sugar-free snacks. The American Diabetes Associ-
ation and the American Dietetic Association per-
mit popcorn as a bread exchange on weight-
control diets; the Feingold Diet for hyperactive
children permits popcorn because it contains no
artificial additives. Nutrition experts agree that
popcorn is a tasty, economical and healthy food.
Popcorn is a whole grain and its nutritive
value is retained inside the hull until it is popped.
The US Department of Agriculture has estimated
the nutritive values in popped popcorn
(Table 12.3). The figures may vary because of
the actual weight or size of the kernels. Popcorn
with high-quality carbohydrate and high fibre
content coupled with low calories makes it
148 V.K. Yadav and P. Supriya

Table 12.3 Nutritive value of popcorn 12.5.4 Future Strategies for Popcorn
Weight 0.5 to 1 ounce in India
Calories 50 to 110 calrefers
to unbuttered popcorn To date, no hybrid of popcorn is available in
Carbohydrates 10 to 22 g India. Thus, the produce has less uniformity,
Protein 2 to 4 g which in turn affects its market acceptance, par-
Fat 1 to 2 g ticularly in international market. Therefore, there
Calcium 2 mg
is a need to develop popcorn hybrids in India.
Iron 0.4 to 0.6 mg
A second problem associated with Indian pop-
Niacin 0.2 to 0.6 mg
corn cultivars is less popping ratio. Best popcorn
Source: Rakshit et al. (2003)
variety has popping ratio of 20:1 as against 40:1
in the USA. Thus, there is scope for breeding
popcorn cultivars with enhanced popping ratio
longer to chew. Compared ounce for ounce with
and with soft texture and bigger volume. There
beef, popcorn provides 67 % as much protein,
is need to develop kernels with thick pericarp,
110 % as much iron and an equal amount of
hard coating with soft starch cavity inside. For
calcium. A pint of popcorn contains three times
this purpose, efforts may be made towards
as much phosphorus as a pint of milk. A cup of
screening of germplasm and their utilisation in
unbuttered popcorn contains fewer calories than
breeding.
half a medium grapefruit.
Popping ratio of popcorn is very much depen-
Popcorn is a very good dieting aid. Eaten just
dent on storage as well as popping temperature
before a meal, it will take the edge off an appetite.
and moisture. Therefore, appropriate drying and
The cellulose is excellent dietary roughage and
storage of popcorn grains are essential. In the
compares to bran flakes. Digestible carbohydrates
USA, and other developed countries, food
in the popcorn provide energy. When popcorn is
industries pack popcorn with appropriate oil,
being used as part of a diet programme, the butter
butter and salts in popping bags. These bags are
may be skipped and just light salt with the popped
ready to pop in the microwave. Such packings
kernels may provide a good taste.
are very handy for end users and fetch very good
market price. Such type of entrepreneurship is
much needed in India. This will be possible only
12.5.3 Value-Added Products with support from the food industry and policy
of Popcorn makers (Rakshit et al. 2003).

A number of value-added products of popcorn

have been developed. The following are some of
the products developed for commercial use: 12.6 Industrial Products of Corn
Apple popcorn brittle
Ballpark popcorn crunch Corn is used in more than 3,000 products which
Beach party popcorn include adhesives, antibiotics, automobiles, baby
Boston tea party popcorn food, breakfast cereals, canned vegetables,
Caramel corn crunch cheese spreads, chocolate products, printings,
Caramel nut popcorn crunch cosmetics, crayon and chalk, dessert powders,
Cherry almond popcorn clusters dyes, edible oil, finished leather, insecticides,
Chilli corn ketchup, livestock feed, malted products, paper
Red cinnamon popcorn manufacturing, pharmaceuticals, drugs and
Swiss onion popcorn carpets, shoe polish, soft drinks, textiles, wheat
Pina colada popcorn bread, yogurts, etc. (Fig. 12.4). Corn is subjected
Patchwork popcorn party mix to wet and dry milling to separate the grain into its
Kettle corn components, which are used as food ingredients.
12 Value Addition in Maize
Fig. 12.4 Various industrial products developed from corn
12.6.1 Dry Milling
Two different approaches have been used for dry
milling of corn: degerming and non-degerming.
Older systems did not remove the germ before
grinding the corn into grits, meal and flour, and
the resulting products had short shelf lives, being
prone to developing rancid flavours and odours.
Today, no more than 20 % of corn is milled by
this traditional or unbolted dry milling process.
Early systems used stone mills for particle reduc-
tion and sieves to separate products on the basis
of particle size.
12.8
12.6.2 Wet Milling
The dry milling industry could not provide starch
(A) Starch: Two-thirds of the starch produced by
of sufficient purity to process into good syrups.
The wet milling industry has grown rapidly in
recent years due to demand for high-fructose
corn syrup (to replace largely imported cane
sugar used in soft drinks) and fuel ethanol (to
increase octane of unleaded fuels, extend petro-
leum reserves and improve air quality).
12.7 Use of Corn in Feed and Food
Corn as livestock feed: Corn is the most
widely used feed stuff for rearing poultry,
piggery and livestock because of its abundant,
predictable and widespread supply; low cost;
high digestibility; and feed conversion with
minimal processing. Corn rarely contains
any toxins, such as the microbial toxins afla-
toxin and fumonisin.
149
Corn as food: Corn may be converted with
little processing into common foods in the
American diet. Alternatively, dry and wet
milling have evolved to take corn apart into
its various components and convert them into
a wide range of food ingredients used in
todays highly engineered and processed
foods. These ingredients are becoming more
important as consumers demand processed
and convenient foods with long shelf life. In
Latin America, Africa, and Asia, it is a staple
food comprising a high proportion of diets.
Use of Corn in Developing
Intermediary Raw Products
wet mills is used for industrial products in
the USA, such as adhesives in paper and
building materials, paper coatings and
sizing, textile sizing, charcoal binders, and
recently in degradable plastics. Both native
and modified starch are used depending
upon the functional properties required.
The starch include the following:
(a) Maize starch: Maize starch exhibits
all the properties of native starch. It
improves efficiency in weaving, there is
no danger of thinning down, and no
foaming during cooking.
(b) Dextrin: It is of two typeswhite and
yellow. White dextrin is an intermediate
product between starch and sugars
derived from starch. Yellow dextrin is
soluble gummy carbohydrate formed
by partial hydrolysis of starch. Yellow
150
dextrin or canary dextrins, as they are
frequently called, are made by roasting
starch with less acid and more heat.
(c) Synthosize: Synthosize is a versatile
product for sizing of blended synthetic
yarn in modern textile industry. It not
only helps in reducing droppings and
breakages but also increases loom effi-
ciency to a great extent, which is the
demand of every modern textile unit.
The use of synthosize completely
eliminates the addition of PVA in size
mix for sizing of blended yarn.
(d) Papyrox: Papyrox is extremely white,
oxidized starch with high brightness. It
is a highly versatile product that finds
wide applications in paper, textile, adhe-
sive, building material and other such
industries.
(e) Thintrite-90: Thintrite-90 is a starch
material in the form of superficially
unchanged granules obtained by action
of chemicals on starch.
(B) Specialty chemicals by chemical processes:
In addition to modifying the starch, syrups
can be converted by chemical processes into
many specialty chemicals including organic
acids, polyols, glucosides and glucose esters.
For many years, corn cobs have been
converted into furfural derivatives.
(C) Ethanol: Corn starch is an economical fer-
mentation feedstock to produce organic
chemicals. The most notable example is
fuel ethanol. Although beverage alcohol
has been produced by corn fermentation for
many years, expansion has been motivated
to extend petroleum reserves and motor
fuels, especially during the early 1980s
when the Organization of Petroleum
Exporting Countries (OPEC) drastically
reduced petroleum supplies and increased
prices.
(D) Specialty chemicals by fermentation: Corn-
derived syrups are easily fermented into a
wide array of specialty chemicals including
organic acids (acetic, citric, gluconic, malic,
succinic, fumaric, propionic, and butyric),
V.K. Yadav and P. Supriya
alcohols (ethanol, isopropanol, and butanol),
ketones (acetone and glucosone), amino
acids (methionine, lysine, and tryptophan),
nucleotides (guanyl, inosinic, and xanthyl
acids), biopolymers (xanthan, pullulan, algi-
nate, and polyhydroxybutyrate), single-cell
proteins and oils, enzymes, vitamins,
antibiotics, and hormones.
(E) Refined corn oil: Corn oil is one of the most
popular international cooking mediums,
particularly popular in the USA and Europe
due to its unique properties. Corn oil is a
co-product of wet milling of corn. The
crude corn oil is refined to remove free
fatty acids and phospholipids. The product
is decolourised and deodourised under ster-
ile conditions to yield sparkling clear oil.
(F) Sorbitol: Sorbitol is a low-calorie refined
sugar which is synthesized from glucose.
Chemically, it is a hexahydric alcohol and
is also known as D-glucitol. It is widely
found in various types of fruits and berries
such as strawberries, cherries, apples and
pears.
(G) Glucopep (liquid glucose): Liquid glucose
sold under the brand name of Glucopep is a
thick syrup manufactured by hydrolysing
starch with acids/enzymes followed by mul-
tiple stages of refining.
12.8.1 Use of Intermediary
Raw Products of Maize in
Developing Finished Product
of Other Industries
Beer: Beer manufacturing is a process of
treating barley to convert and extract the bar-
ley starch to fermentable sugars using the
amylolytic enzymes present in malt followed
by yeast fermentation. However, demand for
lighter, less filling beer, especially in the
USA, has permitted use of more refined car-
bohydrate sources of two types:
(a) Dry adjuncts, primarily dry milled corn
grits, broken rice, refined corn starch,
and more recently dextrose
12 Value Addition in Maize
(b) Liquid adjuncts, namely, corn syrups
Cake mixes: Cake mixes use a pregelatinised
corn starch that forms a paste in cold or warm
water. In baked goods that use yeast for rising,
dextrose is used as a yeast nutrient.
Candies: Corn syrup is used in hard candies to
provide a body, giving them chewiness and
desirable mouthfeel without excessive sweet-
ness. Coated candies use a pyrodextrin corn
starch for the coating.
Carbonated beverages such as Coke: High-
fructose corn syrup (HFCS) blended with
sucrose in a 50/50 blend is sweeter than the
same concentration of sucrose. The use of
HFCS in carbonated beverages is common
throughout Canada and the USA.
Cookies: Corn starch, corn flour, or dextrose
are also used in cookies.
Corn flakes: The flaking grits are cooked to a
rubbery consistency with syrup, malt, salt and
added flavours. After tempering, the cooked
grits are flattened between large steel rolls
followed by toasting in travelling ovens to a
golden brown colour.
Corn meal: Corn meal is a popular dry corn
product because of its long shelf life. It is used
to produce an assortment of chemically leav-
ened bread and fried products like corn bread
and muffins.
Cosmetics: Corn cob when finely ground is
relatively dust-free and very absorbent. This
absorbency makes corn cobs useful carriers
for pesticides, fertilizers, vitamins, hand
soaps, cosmetics, and animal litters.
Granola dips/granola bars: Some types of gra-
nola dips use dextrose as a sweetener.
Gypsum wallboard: Starch containing corn
flour is gelatinised during the manufacturing
process. It functions by controlling the rate of
water loss during drying of the board. Soluble
carbohydrates migrate to the surface and con-
trol the rate of crystallisation of the gypsum,
providing a strong bond between the gypsum
and the liner.
Instant coffee and tea: Maltodextrins are
derived from the wet milling process. They
are a dextrose-equivalent product having
151
complete solubility but little or no sweetness.
Maltodextrins are sprayed on instant tea and
coffee to keep the granules free flowing. This
solution is also used in instant soup mixes or
other packages where the contents must be
kept free flowing.
Mars bar and Twix bar: Many candy bars
contain corn syrup.
Paint and varnish: Tetrahydrofurfuryl alcohol
is a resin developed from processing corn
cobs. These resins are useful in the paint and
varnish industry as solvents for dyes, resins,
and lacquers.
Paper products: Paper products use raw starch
in the manufacturing process. The properties
of high paste viscosity and strong gels are
useful in specially coated papers.
Pyrodextrins are also used for paper
manufacturing for the adhesive property on
remoistenable gums for postage stamps and
packaging tape.
Pharmaceutical such as aspirin: An oxidised
starch paste, which dries to a clear, adherent,
continuous film, is spread in a thin layer over
the aspirin. Some intravenous (IVs) fluids
consist of dextrose and water solutions.
Antibiotics: Preferred carbohydrate sources
are corn syrup, dextrose, corn starch, lactose
and sucrose. Corn steep liquor was early
found to provide a ready source of soluble
nitrogenous nutrients along with unknown
growth factors that stimulate antibiotic pro-
duction. Over 85 different types of antibiotics
are produced using corn.
Snack foodscorn chips and Doritos: These
snack foods are generally made from whole
corn (corn meal). The high starch content of
corn meal and flour is important in giving a
high puff in preparation of extruded (pressed)
snack products in which a delicate corn
flavour is desired.
Spark plugs: Starch is used in the production
of the porcelain part of spark plugs.
Rubber: In the production of tyres, corn
starch is sprinkled on the moulds before tooth-
paste as a low-calorie, water-soluble bulking
agent.
152
Whiskey: The major carbohydrate source in
the production of whiskey is corn. A typical
Canadian whiskey is made from a mixture
of about 90 % corn, 5 % rye and 5 % barley
malt.
Yogurt: Some of the different brands of
yogurt use corn syrup as a sweetener
(Venkatesh et al. 2003)
12.9 Conclusion
The value-added products prepared from maize
are not only nutritious but also easy to prepare.
Keeping in view the nutritional profile of maize,
the development of these products will not only
diversify the uses of maize but also will be
beneficial for human health especially in com-
bating malnutrition. There is a need to develop
and popularize value-added products based on
quality protein maize and baby corn among
housewives so that they include these pre-
parations in their daily diet. The development
of industrial products and dehydration, freezing,
and canning of baby corn can be taken up as
V.K. Yadav and P. Supriya
entrepreneurial activity by the entrepreneurs.
The increased utilisation of maize will encour-
age farmers in improving the production of
maize which may indirectly help in improving
economic standards of farmers. Efforts towards
commercialisation of maize-based value-added
products through self-help groups, food
industries, etc. are also required.
References
CIMMYT (2000) Science and sustenance. CIMMYT,
Mexico, pp 67. ISBN 01889214
Kawatra A, Sehgal S (2007) Value added products of
maize. Report of the national conference on doubling
maize production, IFFCO Foundation, New Delhi,
pp 7685
Prasanna BM, Vasal SM, Kassahun B, Singh NN (2001)
Quality protein maize. Curr Sci 81:13081319
Rakshit S, Venkatesh S, Shekar JC (2003) Pop corn,
Directorate of Maize Research, New Delhi, pp 116
Singh U (2006) Quality protein maize products for human
nutrition. Directorate of Maize Research, New Delhi,
pp 121
Venkatesh S, Kumar RS, Shekar JC (2003) Technologies
for processing speciality maize in India. Directorate
of Maize Research, New Delhi, pp 117
 Fodder Quality of Maize: Its
Preservation 13
D.P. Chaudhary, S.L. Jat, R. Kumar, A. Kumar, and B. Kumar

Abstract
Green fodder is an important component of animal husbandry. The
growth of dairy sector primarily depends upon the availability of
nutritious fodder. Maize is one of the most nutritious non-legume
green fodders. The high acceptability of maize as fodder can be judged
from the fact that it is free from any anti-nutritional components.
Maize is quick growing, yields high biomass, and is highly palatable.
It contains sufficient quantities of protein and minerals and possesses
high digestibility as compared to other non-legume fodders. It contains
high concentrations of soluble sugars in the green stage, which makes
it most fit for preservation as silage. The abundance of green fodder
due to increasing cultivation of specialty corn could greatly help in
boosting the prospects of dairy sector in the peri-urban regions of the
country.

13.1 Introduction production, around 40 kg of green fodder is

The economics of milk production is largely
dependent upon the quality of nutritious fodder
fed to milch animals. Feeding of green forages
compared to concentrates lowers the cost of milk
production substantially. For optimum milk
D.P. Chaudhary (*)  S.L. Jat  R. Kumar
Directorate of Maize Research, New Delhi, India
e-mail: chaudharydp@gmail.com
A. Kumar
Central Soil and Salinity Research Institute, Karnal, India
B. Kumar
Guru Angad Dev Veterinary and Animal Sciences
University, Ludhiana, India
required to feed per animal per day. However, a
huge deficit exits between the demand and sup-
ply of green fodder in India. At present, there
exists around 63 % deficiency of green fodder
and 23.5 % deficiency of dry fodder in India
(Singh 2009). Therefore, to meet the needs of
the ever-increasing livestock population, the pro-
duction as well as productivity of fodder needs to
be increased. The increasing cultivation of
cereal and cash crops, however, contributed
towards a decline in the area under fodder culti-
vation. There is a tremendous pressure of live-
stock on the available total feed and fodder,
as the land available for fodder production has
been decreasing. Moreover, green forages are
rich and the cheapest source of carbohydrates,

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 153
DOI 10.1007/978-81-322-1623-0_13, # Springer India 2014
154 D.P. Chaudhary et al.

protein, vitamins and minerals for dairy animals. Table 13.1 Major legume and non-legume forages of
Therefore, by providing sufficient quantities of India
fodder, instead of costly concentrates, to the S. No. English name Botanical name
milch animals, the cost of milk production can be Legume fodders
considerably reduced. Maize is one of the most 1. Egyptian clover Trifolium alexandrinum
important non-legume green fodders. It is a tall, 2. Persian clover Trifolium resupinatum
leafy plant having biomass yields to the tune of 3. Indian clover Melilotus spp.
400500 q/ha. It is a highly nutritious, palatable 4. Lucerne/alfalfa Medicago sativa
fodder, free from any unwanted anti-quality 5. Cowpea Vigna unguiculata
6. Cluster bean Cyamopsis tetragonoloba
components. Green maize is rich in protein and
Non-legume fodders
possesses sufficient quantities of soluble sugars
1. Maize Zea mays
required for proper ensiling. In this chapter, the
2. Sorghum Sorghum bicolor
fodder quality of maize is discussed in comparison
3. Pearl millet Pennisetum glaucum
to other non-legume forages along with 4. Teosinte Zea mexicana
the methodology for its preservation as silage. 5. Guinea grass Panicum maximum
6. Oat Avena sativa
7. Ryegrass Lolium spp.
13.2 Green Fodders

Green fodder production is the most challenging

activity in the dairy enterprise. It not only reduces
most important food crop after rice and wheat. It
the feeding cost but also keeps the animals
is the crop with the highest per day productivity.
healthy, reduces micronutrient deficiencies and
The production as well as productivity of maize
increases milk production. Forages usually con-
is growing in India. In the XI five-year plan,
tain relatively large concentrations of cellulose,
maize achieved the highest growth rate (6.7 %)
hemicelluloses, and lignin, as well as variable
among cereals as against the required growth rate
amounts of non-fibrous carbohydrates and
of 4.7 % set by the planning commission of India.
proteins. Dairy animals obtain nutrients, viz.,
It is an excellent crop in terms of biomass pro-
energy, protein, fibre, minerals, vitamins and
duction also. Since the production as well as
water from the forages for the maintenance of
productivity of maize is increasing, the availabil-
their body and optimum performance. There are
ity of biomass from maize is also increasing by
many legume and non-legume fodder varieties
the same magnitude. Maize straw has been used
available for cultivation. Ideal characteristics of
as animal fodder since long. The fodder quality
good fodder varieties are short duration, high
of green maize is considered best among non-
biomass potential, nutritious and tasty fodder,
legume forage crops. Maize is considered ideal
suitability for preservation and negligible
forage because it grows quickly, produces high
concentrations of anti-nutritional components.
yields, is palatable, is rich in nutrients, and helps
Green fodders can be broadly divided into two
to increase body weight and milk quality in cattle
categories, viz., legumes and non-legumes. Some
(Sattar et al. 1994). As fodder for livestock,
of the common green forages cultivated in India
maize is excellent, highly nutritive and sustain-
are listed in Table 13.1.
able (Hukkeri et al. 1977, Iqbal et al. 2006). It is
commonly grown as a kharif fodder in the north-
13.3 Maize as Fodder western regions of India. Its quality is much
better than sorghum and pearl millet, since both
Maize (Zea mays) is one of the most important sorghum as well as pearl millet possess anti-
crops having wider adaptability to varied quality components such as hydrocyanic acid
agroclimatic conditions. In India, it is the third and oxalate, respectively.
13 Fodder Quality of Maize: Its Preservation 155

Table 13.2 Comparative nutritional quality of non-legume fodders

Harvesting stage In-vitro dry matter
Days after Crude Protein digestibility
Fodder crop Physiological stage sowing (DAS) (CP) (%) (IVDMD) (%)
Maize Silk to milk stage 5565 118 6852
Bajra Boot stage 4555 107 6255
Sorghum Initiation of flowering 7080 87 6057
Teosinte Pre-flowering 8085 97 6258
Sudax Subsequent cutting after 30 days 6570 117 6055
Napier bajra One meter height and Subsequent 5560 117 6055
hybrid cutting after 30 days
Guinea grass One meter height and subsequent cutting 5560 108 6057
after 2530 days
Source: Gupta et al. (2004)

13.4.1 Fodder Quality of Maize

13.4 Forage Quality
Forage quality is defined in various ways but is
often poorly understood. Though important, for-
age quality often receives far less consideration
than it deserves. Forage quality can be defined as
the extent to which forage has the potential to
produce a desired animal response. Adequate
animal nutrition is essential for high rates of
gain, ample milk production, efficient reproduc-
tion and adequate profits. However, forage qual-
ity varies greatly among and within forage crops.
Analysing forages for nutrient content can be
useful in determining whether quality is adequate
and to guide proper ration supplementation. In
recent years, advances in plant and animal breed-
ing, introduction of new products and develop-
ment of new management approaches have made
it possible to increase animal performance. How-
ever, for this to be realised, there must be addi-
tional focus on forage quality. Many factors
influence forage quality. Some of these include
palatability (whether the animals eat the forage),
intake (how much the animal will eat), digestibil-
ity (the extent to which forage is absorbed as it
passes through an animals digestive tract), nutrient
content, anti-quality components (tannins, nitrates,
alkaloids, cyanoglycosides, oxalates, estrogens
and mycotoxins), and lastly animal performance,
which is the ultimate test of forage quality.
Desirable forage characteristics include high dry
matter yield, high protein concentration, high
energy concentration (high digestibility), high
intake potential (low fibre content), and optimum
dry matter concentration at harvest for accept-
able forage fermentation (Carter et al. 1991).
Maize is an ideal forage crop as it is quick grow-
ing, high yielding, palatable and nutritious.
Among the cultivated non-legume fodders,
maize is the most important crop that can be
grown round the year under irrigated conditions.
It is free from any anti-nutritional components
and is considered a valuable fodder crop. It
contains high concentrations of protein and
minerals and possesses high digestibility. Two
maize varieties, namely, J-1006 and African
tall, are developed and released for commercial
cultivation of fodder in India. The nutritional
quality of maize as compared with other non-
legume fodders is expressed in Table 13.2.
13.5 Specialty Maize as Fodder
Apart from grain, maize is also grown for some
special uses such as baby corn, sweet corn and
popcorn. Maize cultivated for this purpose is
called specialty maize. Specialty corn (baby
corn and sweet corn) cultivation is gradually
156 D.P. Chaudhary et al.

Table 13.3 Green fodder yield, dry matter, crude protein, and crude fibre content of baby corn genotypes
GFY (t/ DM (%) I DM (%) II CP (%) I CP (%) II CF (%) I CF (%) II
Genotype Use ha) harvest harvest harvest harvest harvest harvest
JH-3459 Baby 38.12 21.19 21.13 8.72 7.43 24.77 28.67
Parkash corn 30.14 20.55 20.91 7.00 6.70 26.80 28.77
PMH-2 40.14 21.22 22.47 8.46 8.31 26.33 23.13
J-1006 Fodder 46.67 24.28 24.69 7.44 5.83 26.13 23.87
African 30.99 22.24 22.61 7.14 5.54 30.20 28.67
tall
Source: Chaudhary et al. (2012)

Table 13.4 Neutral detergent fibre, acid detergent fibre, total ash, and in vitro dry matter digestibility of baby corn
genotypes
IVDMD IVDMD
NDF (%) NDF (%) ADF (%) ADF (%) Ash (%) Ash (%) (%) I (%) II
Genotype Use I harvest II harvest I harvest II harvest I harvest II harvest harvest harvest
JH-3459 Baby corn 64.13 66.63 40.80 43.13 8.00 6.23 58.23 55.90
Parkash 64.97 64.67 42.87 44.73 7.33 6.70 63.73 61.80
PMH-2 65.93 64.67 46.77 45.83 6.60 6.10 64.33 58.60
J-1006 Fodder 69.87 72.80 40.37 45.47 7.27 7.43 58.07 52.86
African 67.57 66.10 38.27 38.73 5.97 6.60 65.00 57.63
tall
Source: Chaudhary et al. (2012)

increasing, particularly in the peri-urban regions
of the country. Baby corn is a very delicious and
nutritious vegetable and has been considered to
be a high-value agriculture produce in national
and international markets. The export potential
of baby corn provides further boost to its cultiva-
tion. On the other hand, sweet corn is being
utilised in hotels and big restaurants. Many
value-added products made from baby corn and
sweet corn are being used for human consump-
tion. A significant quantum of green biomass is
available from specialty corn cultivation, which
can efficiently be used as animal fodder. Yield
potential and nutritional quality of some
promising genotypes such as JH-3459, Parkash,
and PMH-2 grown for baby corn along with two
most common fodder maize varieties, viz., Afri-
can Tall and J-1006, were evaluated. Green
samples were taken at I as well as II harvest of
baby corn. After harvesting the baby corn, the
stalk was used for forage quality analysis. The
samples were analysed for various parameters of
fodder quality such as green fodder yield (GFY);
dry matter (DM); crude protein (CP); fibre
components, viz., crude fibre (CF), neutral deter-
gent fibre (NDF), and acid detergent fibre (ADF);
in vitro dry matter digestibility (IVDMD); and
total ash (TA) (Tables 13.3 and 13.4).
The data show that the nutritional quality of baby
corn stalks is almost at par with the maize grown for
fodder purpose. Although biomass from baby
cornstalks was little less compared to fodder maize
J-1006 and African tall, there is little difference in
terms of crude protein and in vitro dry matter digest-
ibility. The woodiness is also comparable. It means
the baby corn is as good as fodder maize.
13.6 Ensiling
Forage conservation is a key element for produc-
tive and efficient ruminant livestock farms. It
permits a better supply of quality feed when
forage production is low or dormant. Forage
conservation also provides farmers with a
means of preserving forage when production is
13 Fodder Quality of Maize: Its Preservation
faster than its adequate utilisation by grazing
animals. This prevents lush growth from becom-
ing too mature. Consequently, forage conserva-
tion provides a more uniform level of high-
quality forage for ruminants. Ensilage has many
advantages over the other methods for preserva-
tion of nutrients, particularly from forages.
Silage is the material produced by controlled
fermentation of nutrients under an anaerobic
condition. The fermentation process is
governed by micro-organisms present in fresh
herbage to maintain anaerobic conditions and
discourage clostridial growth with minimum
loss of nutrients.
13.6.1 Ensiling of Maize
Maize is an excellent crop for ensiling. It
possesses high energy value. For proper fermen-
tation, the crop might possess sufficient
quantities of moisture as well as soluble
carbohydrates which are converted to lactic acid
during the process of fermentation. Maize
cultivated for green fodder and baby corn pur-
pose possesses the required moisture and solu-
ble sugars and, therefore, is most suitable for
ensiling. Maize silage is becoming more impor-
tant in dairy rations. Maize is valued because of
its high yield and ability to make excellent
silage, and it can be harvested in a single oper-
ation without significant leaf loss. Cows fed
corn silage produced more milk and consumed
more silage dry matter in both trials than those
fed sorghum silage (Lance et al. 1964). Corn
silage is used extensively for lactating dairy
cows that require high-energy feed for maxi-
mum milk production (Marsalis et al. 2010;
Irlbeck et al. 1993).
13.6.2 Methodology of Ensiling
The quality of silage depends on the stage at
which the fodder is harvested. Maize is best
suited to be ensiled when the grains are in the
milking stage. In the tropics this is usually found
in 6070 days after sowing. The dry matter
157
content of whole plant should be around
2530 %. Maize is ready for ensiling if the dry
matter in the grain has reached a value between
56 % and 60 %. Harvesting at this recommended
time will ensure optimum compaction properties,
reduced tendency to heating up, and mould for-
mation. Silage making is a simple process which
can be carried out manually in the farm area by
employing a few labourers. The first and fore-
most thing in silage making is the digging of a
pit. A rectangular pit is to be dug near the cattle
shed whose size depends upon the number of
animals along with the availability of fodder. If
the fodder is sufficiently available, then the time
duration of feeding could be considered in decid-
ing the size of the pit. Usually one cubic meter pit
can accommodate roughly 56 quintals of green
fodder. Crop should be chaffed to 57 cm length
before ensiling. For good silage, the chop length
should be kept shorter. Chaffed silage is more
palatable to livestock and has little chance of
secondary fermentation. The next step is filling
the pit with chaffed fodder. For this purpose
fodder is spread up to a height of 1 ft in the pit
followed by compression. This process is
repeated till the pit is filled with fodder. The
major precaution during this process is to
exclude as much air as possible from the chaffed
fodder by compressing it properly. This is
executed by pressing the material through man-
ual labour or mechanically by using a tractor.
Care should be taken that material on the sides
and edges are properly compressed. Raise the
fodder heap above the ground level up to a height
of around one meter. Finally, add some more
fodder in the central portion of the heap and
then trample it. Packing is important to create
anaerobic conditions. It should be thoroughly
pressed so that no air pocket is left in the silo;
otherwise chances of mould formation will be
there which will spoil the silage. After filling,
silo should be covered with polythene sheet
followed by a layer of soil, etc. Some cracks
may develop in the covered soil over time.
These are to be plugged immediately (Figs.
13.1, 13.2 and 13.3).
After 45 days of ensilage, the silage will be
ready to use and, therefore, can be removed for
158
Fig. 13.1 Filling of pit
Fig 13.2 Huge stock of
silage
feeding. Care should be taken in removing the
silage from the pit. It should not be opened from
one side only. Cover should be kept firmly in
place as long as possible and the minimum face
should be exposed at one time. The sugars,
proteins, and lactic acid present in the silage are
subject to attack by mould growth and oxidation
as some air is allowed for fermentation and
causes loss of feeding value and intake by the
animals.
D.P. Chaudhary et al.
13.6.3 Silage Quality
Silage quality is determined mainly by the phys-
ical state, that is, colour and odour. Good quality
silage should be light brown in colour and have
the smell of vinegar. Poorly fermented silage
will be dark in colour and foul smelling due to
the production of butyric acid. Chemically,
it should have the following characteristics:
(i) pH below 4.2 (ii) ammoniacal nitrogen of
13 Fodder Quality of Maize: Its Preservation 159

Fig. 13.3 Silage of baby

corn

Table 13.5 Common end products of silage fermentation

Positive or
Item negative Action(s)
pH + Low pH inhibits bacterial activity
Lactic acid + Inhibits bacterial activity by lowering pH
Acetic acid Associated with undesirable fermentations
+ Inhibits yeasts responsible for aerobic spoilage
Butyric acid Associated with protein degradation, toxin formation and large
losses of DM and energy
Ethanol Indicator of undesirable yeast fermentation and high DM losses
Ammonia High levels indicate excessive protein breakdown
Acid detergent insoluble High levels indicate heat-damaged protein and low energy content
nitrogen (ADIN)

total N, less than 10 % of total N; (iii) butyric

acid, less than 0.2 %; and (iv) lactic acid 312 %.
The end products of silage fermentation are often
monitored to assess silage quality, and the com-
position of normal silages is presented in
Table 13.5.
Thus, it could be concluded that maize is an
excellent crop which could effectively be
utilised as animal fodder. Specialty corn is
going to play an important role in the socio-
economic perspective of rural folk as the dual
purposes of agriculture as well as livestock are
fulfilled by its cultivation. Maize silage could
greatly help towards reducing green fodder
scarcity and provide a much desired boost to
the dairy sector.
References
Carter PR, Coors JG, Undersander DJ, Albrecht KA,
Shaver RD (1991) Corn hybrids for silage: an update.
Roth. In: Proceeding. of annual corn & Sorghum
research conference, 46th. Chicago. American Seed
Trade Association, Washington DC. pp 141164
Chaudhary DP, Kumar A, Mandhania SS, Srivastava P,
Kumar RS (2012) Maize as fodder? An alternate
approach. Technical Bulletin, Directorate of Maize
Research, Indian Council of Agricultural Research,
New Delhi
Gupta BK, Bhardwaj BL, Ahuja AK (2004) Nutritional
value of forage crops of Punjab. Punjab Agricultural
University Publication, Ludhiana
Hukkeri SB, Shukla NP, Rajput RK (1977) Effect of levels
of soil moisture and nitrogen on the fodder yield of oat
on two types of soils. Indian J Agron 47:204209
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Iqbal A, Ayub M, Zaman H, Ahmed R (2006) Impact of
nutrient management and legume association on agro-
qualitative traits of maize forage. Pak J Bot 38:10791084
Irlbeck NA, Russell JR, Hallauer AR, Buxton DR (1993)
Nutritive value and ensiling characteristics of maize
stover as influenced by hybrid maturity and genera-
tion, plant density and harvest date. Anim Feed Sci
Technol 41:5164
Lance RD, Foss DC, Krueger CR, Baumgardt BR,
Niedermeier RP (1964) Evaluation of corn and sor-
ghum silages on the basis of milk production and
digestibility. J Dairy Sci 47:254257
D.P. Chaudhary et al.
Marsalis MA, Angadi SV, Contreras-Govea FE (2010)
Dry matter yield and nutritive value of corn, forage
sorghum, and BMR forage sorghum at different plant
populations and nitrogen rates. Field Crop Res
116:5257
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 About the Editors

Dr. Dharam Paul Chaudhary is working as Senior Scientist (Biochemistry)

at the Directorate of Maize Research (DMR), an institute of the Indian Council
of Agricultural Research (ICAR), New Delhi, which is a leading national
institute looking after various scientific and coordinating activities related to
the development of maize. After completing his studies from Panjab University,
Chandigarh, he served as Assistant Professor at Punjab Agricultural University,
Ludhiana, which is one of the best agricultural universities of India. His present
work is focused towards the development of quality protein maize (QPM),
which is nutritionally superior compared to normal maize. He has good experi-
ence of teaching various graduate as well as postgraduate courses at Punjab
Agricultural University, Ludhiana, and at the Indian Agricultural Research
Institute (IARI), Pusa Campus, New Delhi. He has to his credit more than 30
scientific publications in journals of national and international repute.
Dr. Sandeep Kumar is working as Senior Scientist (Biochemistry) at the
Germplasm Evaluation Division, National Bureau of Plant Genetic
Resources (NBPGR), an institute of the Indian Council of Agricultural
Research (ICAR), New Delhi. He has also worked as Scientist (Biochemis-
try) at the Directorate of Rapeseed-Mustard Research, Sewar, Bharatpur,
which is also an institute of the Indian Council of Agricultural Research
(ICAR). He earned his masters and doctoral degrees in Biochemistry from
Chaudhary Charan Singh Haryana Agricultural University, Hisar, Haryana.
He has good experience of teaching various graduate as well as postgraduate
courses at SBS PGI, Balawala, Dehradun and graduate courses at D. N.
College, Hisar. He has good experience of working on different biochemical
aspects in various groups of crops including cereals, oilseeds and forages.
Presently, he is looking after nutritional and other biochemical aspects of
cereals and oilseeds. He has to his credit more than 20 scientific publications
in journals of national and international repute.
Ms. Sapna Langyan is working as Scientist (Biochemistry) at the Director-
ate of Maize Research, New Delhi. She is engaged in maize quality studies
with special reference to shelf life studies of maize carotenoids. She earned
her Masters degree in Biochemistry from Chaudhary Charan Singh Haryana
Agricultural University, Hisar and M. Phil. from Global Open University,
Nagaland. She is a member of a number of scientific societies and has
published peer-reviewed research papers.

D.P. Chaudhary et al. (eds.), Maize: Nutrition Dynamics and Novel Uses, 161
DOI 10.1007/978-81-322-1623-0, # Springer India 2014